Identificación y Caracterización de Parasporinas, Obtenidas de Aislados de Bacillus spp Nativos de Suelos Colombianos
Digital
- Autores:
-
Castro Pinzón, Yaren Yorlady
- Tipo de recurso:
- Trabajo de grado de pregrado
- Fecha de publicación:
- 2021
- Institución:
- Universidad de Santander
- Repositorio:
- Repositorio Universidad de Santander
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.udes.edu.co:001/5520
- Acceso en línea:
- https://repositorio.udes.edu.co/handle/001/5520
- Palabra clave:
- Bacillus thuringiensis
Parasporins
PCR
Parasporinas
Colombia
Reacción en Cadena de Polimerasa
- Rights
- openAccess
- License
- Derechos Reservados - Universidad de Santander, 2021
| id |
RUDES2_44f160acb25de4746ae7e7271a693e75 |
|---|---|
| oai_identifier_str |
oai:repositorio.udes.edu.co:001/5520 |
| network_acronym_str |
RUDES2 |
| network_name_str |
Repositorio Universidad de Santander |
| repository_id_str |
|
| dc.title.spa.fl_str_mv |
Identificación y Caracterización de Parasporinas, Obtenidas de Aislados de Bacillus spp Nativos de Suelos Colombianos |
| title |
Identificación y Caracterización de Parasporinas, Obtenidas de Aislados de Bacillus spp Nativos de Suelos Colombianos |
| spellingShingle |
Identificación y Caracterización de Parasporinas, Obtenidas de Aislados de Bacillus spp Nativos de Suelos Colombianos Bacillus thuringiensis Parasporins PCR Parasporinas Colombia Reacción en Cadena de Polimerasa |
| title_short |
Identificación y Caracterización de Parasporinas, Obtenidas de Aislados de Bacillus spp Nativos de Suelos Colombianos |
| title_full |
Identificación y Caracterización de Parasporinas, Obtenidas de Aislados de Bacillus spp Nativos de Suelos Colombianos |
| title_fullStr |
Identificación y Caracterización de Parasporinas, Obtenidas de Aislados de Bacillus spp Nativos de Suelos Colombianos |
| title_full_unstemmed |
Identificación y Caracterización de Parasporinas, Obtenidas de Aislados de Bacillus spp Nativos de Suelos Colombianos |
| title_sort |
Identificación y Caracterización de Parasporinas, Obtenidas de Aislados de Bacillus spp Nativos de Suelos Colombianos |
| dc.creator.fl_str_mv |
Castro Pinzón, Yaren Yorlady |
| dc.contributor.advisor.none.fl_str_mv |
Suárez Barrera, Miguel Orlando Rueda Forero, Nohora Juliana |
| dc.contributor.author.none.fl_str_mv |
Castro Pinzón, Yaren Yorlady |
| dc.subject.proposal.eng.fl_str_mv |
Bacillus thuringiensis Parasporins PCR |
| topic |
Bacillus thuringiensis Parasporins PCR Parasporinas Colombia Reacción en Cadena de Polimerasa |
| dc.subject.proposal.spa.fl_str_mv |
Parasporinas Colombia Reacción en Cadena de Polimerasa |
| description |
Digital |
| publishDate |
2021 |
| dc.date.accessioned.none.fl_str_mv |
2021-08-20T13:48:45Z |
| dc.date.available.none.fl_str_mv |
2021-08-20T13:48:45Z |
| dc.date.issued.none.fl_str_mv |
2021-01-28 |
| dc.type.spa.fl_str_mv |
Trabajo de grado - Pregrado |
| dc.type.coarversion.fl_str_mv |
http://purl.org/coar/version/c_71e4c1898caa6e32 |
| dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_7a1f |
| dc.type.content.spa.fl_str_mv |
Text |
| dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
| dc.type.redcol.spa.fl_str_mv |
https://purl.org/redcol/resource_type/TP |
| format |
http://purl.org/coar/resource_type/c_7a1f |
| dc.identifier.local.none.fl_str_mv |
T 33.21 C188i |
| dc.identifier.uri.none.fl_str_mv |
https://repositorio.udes.edu.co/handle/001/5520 |
| identifier_str_mv |
T 33.21 C188i |
| url |
https://repositorio.udes.edu.co/handle/001/5520 |
| dc.language.iso.spa.fl_str_mv |
spa |
| language |
spa |
| dc.rights.spa.fl_str_mv |
Derechos Reservados - Universidad de Santander, 2021 |
| dc.rights.coar.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
| dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
| dc.rights.creativecommons.spa.fl_str_mv |
Atribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0) |
| dc.rights.uri.spa.fl_str_mv |
https://creativecommons.org/licenses/by-nc-nd/4.0/ |
| rights_invalid_str_mv |
Derechos Reservados - Universidad de Santander, 2021 Atribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0) https://creativecommons.org/licenses/by-nc-nd/4.0/ http://purl.org/coar/access_right/c_abf2 |
| eu_rights_str_mv |
openAccess |
| dc.format.extent.spa.fl_str_mv |
85 p |
| dc.format.mimetype.spa.fl_str_mv |
application/pdf |
| dc.publisher.spa.fl_str_mv |
Bucaramanga : Universidad de Santander, 2021 |
| dc.publisher.faculty.spa.fl_str_mv |
Facultad de Ciencias Exactas, Naturales y Agropecuarias |
| dc.publisher.place.spa.fl_str_mv |
Bucaramanga, Colombia |
| dc.publisher.program.spa.fl_str_mv |
Microbiología Industrial |
| institution |
Universidad de Santander |
| bitstream.url.fl_str_mv |
https://repositorio.udes.edu.co/bitstreams/9692b120-3a8d-4d0e-9df6-8e6d786ecee4/download https://repositorio.udes.edu.co/bitstreams/ecc4de6f-565b-4e72-b7d4-ace1aaa19cd1/download https://repositorio.udes.edu.co/bitstreams/cb417a64-8444-43d3-a737-0a0625ddd0e8/download https://repositorio.udes.edu.co/bitstreams/ee92dcc3-7c7a-4b6c-a856-6658ba9c5e8d/download |
| bitstream.checksum.fl_str_mv |
fb9bc65644b5e76bd994bad9ce5e9596 38d94cf55aa1bf2dac1a736ac45c881c c4e3eb32fea9b8227436b597a2240c8d ec5cd2a8ff30fc813f8e6a39a5d08679 |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 |
| repository.name.fl_str_mv |
Repositorio Universidad de Santander |
| repository.mail.fl_str_mv |
soporte@metabiblioteca.com |
| _version_ |
1814158539348770816 |
| spelling |
Suárez Barrera, Miguel Orlando335becb1-2371-418a-acf8-21265141ac00-1Rueda Forero, Nohora Julianafdefebdb-5a74-4c59-ba33-4ff4cc09bca4-1Castro Pinzón, Yaren Yorlady66e85b8e-bf88-4e6c-aed9-ca14c978497a-12021-08-20T13:48:45Z2021-08-20T13:48:45Z2021-01-28DigitalA pesar del desarrollo de diversos procedimientos médicos enfocados hacia el tratamiento del cáncer, al día de hoy siguen existiendo falencias en la preservación de tejidos sanos. Por lo anterior, las nuevas estrategias se han centrado en el uso de moléculas biológicas específicas con potencial anticancerígeno. En este sentido, durante los últimos años, han aumentado las investigaciones con Bacillus thuringiensis (Bt), un microorganismo Grampositivo caracterizado por la producción de inclusiones cristalinas que resultan tóxicas hacia varios órdenes de insectos. En los estudios más recientes se ha reportado que Bt también tiene la capacidad de sintetizar proteínas no insecticidas conocidas como Parasporinas (PS), las cuales producen cristales que presentan citotoxicidad contra diferentes líneas celulares de cáncer. El presente proyecto se enfocó en la identificación de genes parasporales en 13 aislamientos de Bacillus spp de la colección del Laboratorio de Biología Molecular y Biotecnología de la Universidad de Santander aisladas de suelos colombianos, por medio de la estandarización de la técnica molecular de reacción en cadena de la polimerasa (PCR). Los análisis en geles de agarosa permitieron determinar la presencia de amplicones de interés para PS1 en los aislados 87 y 88B y para PS2 en el aislado 67. La comparación de la secuencia de aminoácidos del producto de PCR de PS2 con la proteína de referencia PS2Aa1 mostró un porcentaje de identidad del 99%. Así mismo, se evidenció una relación filogenética estrecha entre las secuencias de los amplicones de interés de los aislados 67 y 88B, sugiriendo un alto nivel de conservación entre los genes que codifican PS1 y PS2. Hasta el momento, este es el primer informe de investigaciones con toxinas PS en el territorio colombiano, indicando la existencia de aislados nativos en el país con potencial para la producción de estas proteínas.Despite the development of various medical procedures focused on the treatment of cancer, to this day there’re still shortcomings in the preservation of healthy tissues. Therefore, new strategies have focused on the use of specific biological molecules with anticancer potential. In this sense, in recent years, research has increased with Bacillus thuringiensis (Bt), a gram-positive microorganism characterized by the production of crystalline inclusions that are toxic towards various orders of insects. In the most recent studies, it has been reported that Bt also has the ability to synthesize non-insecticidal proteins known as Parasporins (PS), which produce crystals that exhibit cytotoxicity against different cancer cell lines. Recently, 6 families of PS have been identified, based on the homology of their amino acid sequences. This project focused on the identification of parasporal genes in 13 strains of Bacillus spp from the collection of the Molecular Biology and Biotechnology Laboratory of the University of Santander isolated from Colombian soils, through the standardization of polymerase chain reaction technique. (PCR). The analysis in agarose gels made it possible to determine the presence of amplicons of interest for PS1 in isolates 87 and 88B and for PS2 in isolate 67. The analysis of the protein profile using SDS-Page of strain 67 showed the presence of a band 37 kDa, corresponding to the weight of PS2. Similarly, the comparison of the amino acid sequence of this strain with the reference protein PS2Aa1 showed a percentage of identity of 99%. So far, this is the first research report with PS toxins in the Colombian territory, indicating that there’re native isolates in the country with potential for the production of these proteins.PregradoMicrobiólogo Industrial1 ed.Resumen ............................................................................................................................ 12 Abstract ............................................................................................................................. 14 Introducción ...................................................................................................................... 16 1. Objetivos ...................................................................................................................... 20 1.1 Objetivo General .......................................................................................................... 20 1.2 Objetivos Específicos ................................................................................................... 20 2. Pregunta de Investigación ............................................................................................ 21 3. Hipótesis de Investigación............................................................................................ 22 3.1 Hipótesis Nula .............................................................................................................. 22 3.2 Hipótesis Alternativa .................................................................................................... 22 4. Marco Teórico .............................................................................................................. 23 4.1 Bacillus thuringiensis ................................................................................................... 23 4.2 Proteínas Cry ................................................................................................................ 24 4.3 Parasporinas.................................................................................................................. 27 4.3.1 Parasporina 1 (PS1) ...................................................................................................... 29 4.3.2 Parasporina 2 (PS2) ...................................................................................................... 32 4.3.3 Parasporina 3 (PS3) ...................................................................................................... 33 4.3.4 Parasporina 4 (PS4) ...................................................................................................... 35 4.3.5 Parasporina 5 (PS5) ...................................................................................................... 35 4.3.6 Parasporina 6 (PS6) ...................................................................................................... 36 5. Estado del Arte ............................................................................................................. 38 6. Marco Legal ................................................................................................................. 44 7. Materiales y Métodos ................................................................................................... 46 7.1 Diseño del Estudio........................................................................................................ 46 7.2 Metodología.................................................................................................................. 46 7.2.1 Aislamientos Bacterianos y Condiciones de Cultivo ................................................... 46 7.2.2 Caracterización Macroscópica y Microscópica ............................................................ 46 7.2.3 Extracción y Cuantificación de ADN ........................................................................... 47 7.2.4 Cebadores y Condiciones de Amplificación ................................................................ 48 7.2.5 Clonación y Secuenciación Nucleotídica de Amplicones ............................................ 51 7.2.6 Análisis Filogenético .................................................................................................... 52 7.2.7 Extracción Total de Proteínas ....................................................................................... 53 7.2.8 Análisis del Perfil Proteico Mediante SDS-PAGE....................................................... 54 8. Resultados y Discusión ................................................................................................ 55 8.1 Caracterización Macroscópica y Microscópica de Aislamientos de Bacillus spp ....... 55 8.2 Amplificación de Genes ps Mediante PCR .................................................................. 59 8.3 Clonación y Secuenciación Nucleotídica de Amplicones ............................................ 61 8.4 Análisis Filogenético .................................................................................................... 62 8.5 Análisis del Perfil Proteico Mediante SDS-PAGE....................................................... 68 9. Conclusiones ................................................................................................................ 70 Referencias Bibliográficas ................................................................................................ 71 Apéndices .......................................................................................................................... 8485 papplication/pdfT 33.21 C188ihttps://repositorio.udes.edu.co/handle/001/5520spaBucaramanga : Universidad de Santander, 2021Facultad de Ciencias Exactas, Naturales y AgropecuariasBucaramanga, ColombiaMicrobiología IndustrialDerechos Reservados - Universidad de Santander, 2021info:eu-repo/semantics/openAccessAtribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0)https://creativecommons.org/licenses/by-nc-nd/4.0/http://purl.org/coar/access_right/c_abf2Bacillus thuringiensisParasporinsPCRParasporinasColombiaReacción en Cadena de PolimerasaIdentificación y Caracterización de Parasporinas, Obtenidas de Aislados de Bacillus spp Nativos de Suelos ColombianosTrabajo de grado - Pregradohttp://purl.org/coar/resource_type/c_7a1fTextinfo:eu-repo/semantics/bachelorThesishttps://purl.org/redcol/resource_type/TPhttp://purl.org/coar/version/c_71e4c1898caa6e32Todas las AudienciasAbe Y, Shimada H, Kitada S (2008) Raft-targeting and Oligomerization of Parasporin-2, a Bacillus thuringiensis Crystal Protein with Anti-Tumour Activity. Journal of Biochemistry, 143, 269-275.Abe Y, Inoue H, Ashida H, Maeda Y, Kinoshita T, Kitada S. (2017). Glycan region of GPI anchoredprotein is required for cytocidal oligomerization of an anticancer parasporin-2, Cry46Aa1 protein, from Bacillus thuringiensis strain A1547. Journal of Invertebrate Pathology, 142:71-81.Aberkane L, Nacer-Khodja A, Djenane Z, Neila L, Ouafek A, Bousiama L, Grib H, Mameri N, Nateche F, Djefal (2020) In Vitro Cytotoxicity of Parasporins from Native Algerian Bacillus thuringiensis Strains Against Laryngeal and Alveolar Cancers. Current Microbiology 77:405–414Akiba T, Abe Y, Kitada S, Kusaka Y, Ito A, Ichimatsu T, Katayama H, Akao T, Higuchi K, Mizuki E, Ohba M, Kanai R, Harata K (2004) Crystallization of parasporin‐2, a Bacillus thuringiensis crystal protein with selective cytocidal activity against human cells.Akiba T., & Okumura S. (2017). Parasporins 1 and 2: Their structure and activity. Journal of Invertebrate Pathology, 142, 44–49.Aktar N, Karim M, Nargis S, Rahman M, Begum A, Hoq M (2019) In silico Studies of Parasporin Proteins: Structural and Functional Insights and Proposed Cancer Cell Killing Mechanism for Parasporin 5 and 6. Microbial Bioactives 2(1), 082-090.Altschul S, Madden T, Schäffer A, Zhang J, Zhang A, Miller W, Lipman D (1997) Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Res. 25, 3389–3402Ammons D, Short J, Bailey J, Hinojosa G, Tavarez L., Salazar M, Rampersad J (2016). Anti-cancer Parasporin Toxins are Associated with Different Environments: Discovery of Two Novel Parasporin 5-like Genes. Current Microbiology, 72(2), 184–189.Aronson AI, Fritz-James P (1976) Structures and morphogenesis of the bacterial spore coat. Bacteriol Rev 40:360–402Aronson A, Beckman W, Dunn P (1986) Bacillus thuringiensis and related insect pathogens. Microbiol. Rev. 50:1-24.Aston A., Saffie R, Canham L & Ogden J (2005) Nanotechnology Applications for Drug Delivery. Pharm. Tech. Eur. 17 (4), 21–28.Balaraman K (2005) Occurrence and diversity of mosquitocidal strains of Bacillus thuringiensis. J. Vector Borne Dis. 42:81- 86.Beegle C, Yamamoto Y (1992) Invitation paper (C.P. Alexander Fund): History of Bacillus thuringiensis Berliner research and development. Can. Ent. 124: 587-616Becker N (2000) Bacterial control of vector mosquitoes and black flies. In: Entomopathogenic Bacteria: From Laboratory to Field Application. Charles JF, Delécluse A, Nielsen-LeRoux (eds.). Dordrecht, Kluwer Academic Publishers, pp. 383-398.Berliner E (1911) About the sleepiness of the flour moth taup. Z. Gesamte Getreidewe 3: 63-70.Bernhard K, Jarrett P, Meadows M, Butt J, Ellis DJ, Roberts GM, Pauli S, Rodger P, Burges HD (1997) Natural isolates of Bacillus thuringiensis: worldwide distribution, characterization and activity against insect pests. J Invertebr Pathol. 70:59–68Brasseur K, Auger P, Asselin E, Parent S, Côté J, Sirois M (2015) Parasporin-2 from a New Bacillus thuringiensis 4R2 Strain Induces Caspases Activation and Apoptosis in Human Cancer Cells. PLOS One 10(8): 1-22.Bravo A, Gill SS, Soberón M. (2005). Bacillus thuringiensis mechanisms and use. In: Comprehensive molecular insect science. Elsevier B.V. p. 175–206.Bravo A, Soberón M (2007) Las toxinas Cry de Bacillus thuringiensis: modo de acción y consecuencias de su aplicación. Biotecnología 14: 303-313.Bravo A, Gil SS, Soberón M (2007) Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon 49(4): 423-435.Carlson C, Caugant D, Kolsto A (1994). Genotypic Diversity among Bacillus cereus and Bacillus thuringiensis Strains. Appl Environ Microbiol, 60: 1719-1725Carrozi NB, Kramer VC, Warren GW, Evola S, Koziel MG (1991) Prediction of insecticidal activity of Bacillus thuringiensis strains by polymerase chain reaction product profiles. Appl Environ Microbiol. 57:3057–3061Carswell EA, Old LJ, Kassel RL, Green S, Fiore N, Williamson B (1975) An endotoxin-induced serum factor that causes necrosis of tumors. Proc Natl Acad Sci. 72:3666–3670.Chai P, Rathinam X, Hamzah A, Subramaniam S (2016) Characterization of a native Bacillus thuringiensis isolates from Malaysia that produces exosporiumenclosed parasporal inclusion. Emir. J. Food Agric 28(9): 653-659.Chatterjee, S. N.; T. Bhattacharya; T. K. Dangar; G. Chandra (2007) Ecology and Diversity of Bacillus thuringiensis in Soil Environment. African Journal of Biotechnology 6(13):1587-1591.Chubicka T, Girija D, Deepa K, Salini S, Meera N, Chathrattil A, Kunnathully M, Devassy T (2018) A parasporin from Bacillus thuringiensis native to Peninsular India induces apoptosis in cancer cells through intrinsic pathway. J Biosci 43(2): 407–416Clynes R, Towers T, Presta L, Ravetch J (2000). Inhibitory Fc receptors modulate in vitro cytotoxicity against tumor targets. Nature Medicine 6:443–446.Crickmore N, Berry C, Panneerselvam S, Mishra R, Connor T, Bonning B (2020) A structure-based nomenclature for Bacillus thuringiensis and other bacteria-derived pesticidal proteins. Journal of invertebrate pathology. https://doi.org/10.1016/j.jip.2020.107438de La Garza J; Juárez P (2014) El cáncer. (1ª edición). Monterrey, México.Dueñas A, Rojas M, Lucio D, Serrano M, Piñeros M (2015). Investigación en cáncer en Colombia, 2000-2010. Revista Colombiana de Cancerología , 19 (1), 39-46.Ekino K., Okumura S., Ishikawa T., Kitada S., Saitoh H., Akao T., Mizuki, E. (2014). Cloning and Characterization of a Unique Cytotoxic Protein Parasporin-5 Produced by Bacillus thuringiensis A1100 Strain. Toxins, 6(6), 1882–1895.Espino A (2014) Caracterización biológica de parasporinas en cepas nativas de Bacillus thuringiensis. Universidad Autónoma de Nuevo León. 142 p.Fabbri A, Travaglione S, Falzano L, Fiorentini C (2008) Bacterial protein toxins: Current and potential clinical use. Curr Med Chem. 15(11):1116-1125.Federici BA, Lthy P, Ibarra JE (1990) The parasporal body of Bacillus thuringiensis subsp. israelensis: structure, protein composition and toxicity. In: de Barjac H, Sutherland DJ (eds) Bacterial control of mosquitos and blackflies: biochemistry, genetics and applications of Bacillus thuringiensis and Bacillus sphaericus. Rutgers University Press, New Brunswick, pp 16–44Forbes NS (2011) Engineering the perfect (bacterial) cancer therapy. Nature Reviews Cancer: 10: 785-794Glare TR, O'Callaghan M (2000): Bacillus thuringiensis: Biology, Ecology and Safety. Chichester, John Wiley, p. 350.Gonzales E, Granados J, Short J, Ammons D, Rampersad J (2011) Parasporins from a Caribbean Island: Evidence for a Globally Dispersed Bacillus thuringiensis Strain. Curr Microbiol 62:1643–1648.Gordon R, Haynes W, Pang C (1973) The genus Bacillus. En: US Department of Agriculture handbook N° 427. Washington DC., USDA p. 109-26Guerchicoff A (1999) Aspectos básicos y aplicados de la genética molecular de Bacillus thuringiensis. Universidad de Buenos Aires.Hall TA (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT.Nucl. Acids. Symp. Ser. 41:95-98.Harvey A (2008) Natural products in drug discovery. Drug Discovery Today 13:894– 901.Hastowo S, Lay BW, Ohba M (1992) Naturally occurring Bacillus thuringiensis in Indonesia. J Appl Microbiol 73: 108-113.Höfte H, Whiteley H (1989) Insecticidal crystal proteins of Bacillus thuringiensis. Microbiol Rev 53: 242-55.Hu Y (2016) Regulatory Concern of Polymerase Chain Reaction (PCR) Carryover Contamination. DOI: 10.5772 / 66294IARC (2018) Global Cancer Observatory. Obtenido de https://gco.iarc.fr/Ibarra JE, Rincon MC, Orduz S, Noriega D, Benintende G, Monnerat R, Regis L, Claudia MF, de Oliveria M, Lanz H, Rodriguez MH, Sanchez J, Pena G, Bravo A (2003) Diversity of Bacillus thuringiensis strains from Latin America with insecticidal activity against different mosquito species. Appl Environ Microbiol 69:5269–5274Ishiwata S (1901) On a kind of severe flacherie (sotto disease) Dainihon Sanshi Kaiho 114(1):1-5.Ito A, Sasaguri Y, Kitada S, Kusaka Y, Kuwano K, Masutomi K, Ohba M. (2004). A Bacillus thuringiensis crystal protein with selective cytocidal action to human cells. Journal of Biological Chemistry, 279 (20), 21282-21286.Jukes T.H. y Cantor C.R. (1969). Evolución de moléculas de proteínas. En Munro HN, editor, Mammalian Protein Metabolism, págs. 21-132, Academic Press, Nueva York.Jung Y, Mizuki E, Akao T, Côté J (2007). Isolation and characterization of a novel Bacillus thuringiensis strain expressing a novel crystal protein with cytocidal activity against human cancer cells. Journal of Applied Microbiology, 103(1), 65-79.Katayama H, Yokota H, Akao T, Nakamura O, Ohba M, Mekada E, Mizuki E (2005) Parasporin-1, a novel cytotoxic protein to human cells from non-insecticidal parasporal inclusions of Bacillus thuringiensis. J Biochem 137(1):17-25.Katayama H, Kusaka Y, Yokota H, Akao T, Kojima M, Nakamura O, Mekada E, Mizuki E (2007) Parasporin-1, a Novel Cytotoxic Protein from Bacillus thuringiensis, Induces Ca2+ Influx and a Sustained Elevation of the Cytoplasmic Ca2+ Concentration in Toxin-sensitive Cells. Journal of Biological Chemistry 282(10):7742-7752.Katayama, H; Kusaka, Y; & Mizuki, E. (2011).U.S. Patent Application No. 12/935,513. US20090935513. 2009-03-30. Fukuoka Prefectural GovernmentKim HS, Yamashita S, Akao T, Saitoh H, Higuchi K, Park YS, Ohba M (2000) In vitro cytotoxicity of non-Cyt inclusion proteins of a Bacillus thuringiensis isolate against human cells, including cancer cells. Journal of Applied Microbiology 89:16–23.Krishnan V (2013) Investigation of parasporins, the cytotoxic proteins from the bacterium Bacillus thuringiensis. Universidad de Sussex.Kronstad J, Whiteley HR (1986) Three classes of homologous Bacillus thuringiensis crystal-protein genes. Gene 43:29–40Kumar S, Stecher G, Li M, Knyaz C, and Tamura K (2018) MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Molecular Biology and Evolution 35:1547-1549.Lee D-H, Cha IH, Woo DS, Ohba M (2003) Microbial ecology of Bacillus thuringiensis: fecal populations recovered from wildlife in Korea. Can J Microbiol 49: 465-471Lenina N, Naveenkumar A, Sozhavendan E, Balakrishnan N, Balasubramani V, Udayasuriyan V (2014) Characterization of parasporin gene harboring Indian isolates of Bacillus thuringiensis. Biotech 4: 545-551.Liao L, Lin S, Tian L, Chen A, Lin Y (2013) Key aromatic amino acids of anti-hepatoma activity on Parasporin-2. Chinese journal of biotechnology 29(6): 823-835.Liu Y, Lai Q, Goker M, Meier-Kolthoff J, Wang M, Sun Y, Wang L, Shao Z (2015) Genomic insights into the taxonomic status of the Bacillus cereus group. Sci. Rep. 5: 1-11.Lopez J, Ibarra J (1996) Characterization of a novel strain of Bacillus thuringiensis. Appl Environ Microbiol 62:1306–1310López S, Cerón J (2010) Proteínas Cry de Bacillus thuringiensis y su interacción con coleópteros. NOVA 8(14): 121-240.Mansour A (2011) BioEdit: An important software for molecular biology. GERF Bulletin of Biosciences 2(1):60-61Minsalud (2020) Cáncer. Obtenido de https://www.minsalud.gov.co/salud/publica/PENT/Paginas/Prevenciondel-cancer.aspxMizuki E, Ohba M, Akao T, Yamashita S, Saitoh H, Park YS (1999) Unique activity associated with non-insecticidal Bacillus thuringiensis parasporal inclusions: in vitro cell-killing action on human cancer cells. J Appl Microbiol 86:477-486.Mizuki E, Park YS, Saitoh H, Yamashita S, Akao T, Higuchi K, Ohba M (2000) Parasporin, a human leukemic cell-recognizing parasporal protein of Bacillus thuringiensis. Clinical and Diagnostic Laboratory Immunology 7: 625–634.Montero A; Hervás A, Morera R, Sancho S, Córdoba S, Corona J, Rodríguez I, Chajón E, Ramos A (2005) Control de síntomas crónicos: Efectos secundarios del tratamiento con Radioterapia y Quimioterapia. Oncología (Barcelona), 28(3), 41-50.Nagamatsu Y, Okamura S, Saitou H, Akao T, Mizuki E (2010) Three Cry toxins in two types from Bacillus thuringiensis strain M019 preferentially kill human hepatocyte cancer and uterus cervix cancer cells. Biosci Biotechnol Biochem 74:494–498.Ohba M, Aizawa K (1986) Insect toxicity of Bacillus thuringiensis isolated from soils of Japan. J Invertebr Pathol 47: 12-20.Ohba M, Mizuki E, Uemori A (2009) Parasporin, A New Anticancer Protein Group from Bacillus thuringiensis. Anticancer Research 29(1): 427-433.Okumura S, Akao T, Higuchi K, Saitoh H, Mizuki E, Ohba M, Inouye K. (2004). Bacillus thuringiensis serovar shandongiensis strain 89-T-34-22 produces multiple cytotoxic proteins with similar molecular masses against human cancer cells. Letters in Applied Microbiology, 39(1), 89-92.Okumura S, Saitoh H, Wasano N, Katayama H, Higuchi K, Mizuki E, Inouye K (2006) Efficient solubilization, activation, and purification of recombinant Cry45Aa of Bacillus thuringiensis expressed as inclusion bodies in Escherichia coli. Protein Expr. Purif., 47: 144-151.Okumura S, Saitoh H, Ishikawa T, Mizuki E, Inouye, K. (2008). Identification and characterization of a novel cytotoxic protein, parasporin-4, produced by Bacillus thuringiensis A1470 strain. Biotechnology Annual Review, 225–252.Okumura S, Ohba M, Mizuki E, Crickmore N, Côté JC, Nagamatsu Y, Kitada S, Sakai H, Harata K, Shin T (2010). "Parasporin nomenclature" Recuperado [4 de julio de 2020] desde http://parasporin.fitc.pref.fukuoka.jp/Okumura S, Saitoh H, Ishikawa T, Inouye K, Mizuki E (2011) Mode of action of parasporin-4, a cytocidal protein from Bacillus thuringiensis. Biochimica et Biophysica Acta 1808(6): 1476-1482.Organización Mundial de la Salud (2020) World cancer report. Recuperado [23 de octubre de 2020] desde https://www.iarc.fr/cards_page/world-cancer-report/Palma L, Muñoz D, Berry C, Murillo J, Primitivo C (2014) Bacillus thuringiensis Toxins: An Overview of Their Biocidal Activity. Toxins 6: 3296-3325.Patyar S, Joshi R, Prasad D, Prakash A, Medhi B, Das B (2010) Bacteria in cancer therapy: a novel experimental strategy. Journal of Medical Science 17(1):21.Pigott C, King M, Ellar D (2008) Investigating the Properties of Bacillus thuringiensis Cry Proteins with Novel Loop Replacements Created Using Combinatorial Molecular Biology. Applied and Environmental Microbiology 74(11): 3497-3511.Pineda M, Núñez M (2011) Use of bacteria and their products in cancer therapy. Gaceta Mexicana de Oncología 10(6): 366-372Poornima K, Selvanayagam P, Shenbagarathai R (2010) Identification of native Bacillus thuringiensis strain from South India having specific cytocidal activity against cancer cells. Journal of Applied Microbiology 109:348–354Porcar M., Juárez V (2004) Aislamiento y establecimiento de una colección de Bacillus thuringiensis. En Bacillus thuringiensis en el control biológico. Bravo, A. y Cerón, J. eds. Universidad Nacional de Colombia. Bogotá, Colombia. pp. 69-100.Portela D, Chaparro A, López S (2013) La biotecnología de Bacillus thuringiensis en la agricultura. NOVA 11(20): 87-96.Prassad S, Shethna (1974) Purification, crystallization and partial characterization of the antitumour and insecticidal protein subunit from the 5-endotoxin of Bacillus thuringiensis var. thuringiensis. Biochimica et Biophysica Acta, 363: 558--566.Rangel J (2015) La biodiversidad de Colombia: significado y distribución regional. Rev. Acad. Colomb. Cienc. Ex. Fis. Nat. 39(151):176-200Rodríguez J, Martínez L, Cruz N, Cómbita A. (2014). Terapia génica para el tratamiento del cáncer. Rev. colombiana de cancerología. 18(1):27-40.Roh JY, Choi JY, Li MS, Jin BR, Je YH (2017) Bacillus thuringiensis as a specific, safe, and effective tool for insect pest control. International Journal of Microbiology Biotechnology. 17:547.Ruiz de Escudero I., Ibañez I., Padilla M., Carnero A., Caballero P (2004) Aislamiento y caracterización de nuevas aislamientos de Bacillus thuringiensis procedentes de tierras canarias. Bol. San. Veg. Plagas. 30: 703-712.Salas Y, Hernández F, Balagururamy N, De la Fuente N (2019) Biotechnological alternative against cancer: Parasporins of Bacillus thuringiensis. Revista de Ingeniería Biomédica y Biotecnología 3(7): 8-17.Salaverry O (2013) La etimología del cáncer y su curioso curso histórico. Revista Peruana de Medicina Experimental y Salud Publica 30(1), 137-141.Sauka D, Benintende G (2008). Bacillus thuringiensis: Generalidades, un acercamiento a su empleo en el biocontrol de insectos lepidópteros que son plagas agrícolas. Rev. Argentina de Microbiología 40(2): 124-140.Schnepf E, Crickmore N, Van Rie J, Lereclus D, Baum J, Feitelson J (1998) Bacillus thuringiensis and its pesticidal crystal proteins. Microbiol Mol Biol Rev 62: 775-806.Sneath P (1986) Spore forming gram-positive rods and cocci. En: Butler J, editor. Bergey’s Manual of Systematic Bacteriology p. 1104-207.Soberón M, Bravo A (2007) Las toxinas Cry de Bacillus thuringiensis: Modo de acción y consecuencias de su aplicación. Biotecnología 14: 303-313.Suárez M (2018) Estado del arte de parasporinas y su actividad tóxica como agente anticancerígeno bacteriano promisorio. Universidad de Antioquia.Tamura K. (1992). Estimation of the number of nucleotide substitutions when there are strong transition-transversion and G + C-content biases. Molecular Biology and Evolution 9:678-687.Tsuzuki K, Kimura K, Fujii N, Yokosawa N, Indoh T, Murakami T, Oguma K (1990) Cloning and complete nucleotide sequence of the gene for the main component of hemagglutinin produced by Clostridium botulinum type C. Infection and Immunity 58:3173–3177Uemori A, Maeda M, Yasutale K, Ohgushi A, Kagoshima K, Mizuki E, Ohba M (2007) Ubiquity of parasporin-1 producers in Bacillus thuringiensis natural populations of Japan. Naturwissenschaften 94:34–38Uemori A, Ohgushi A, Yasutake K, Maeda M, Mizuki E, Ohba M (2008) Parasporin-1Ab, a Novel Bacillus thuringiensis Cytotoxin Preferentially Active on Human Cancer Cells In Vitro. Anticancer Research 28: 91-96.Velásquez L, Rojas D, Cerón J (2018) Proteínas de Bacillus thuringiensis con actividad citotóxica: Parasporinas. Rev. Colomb. Biotecnol. 20(2): 89-100Verma S, Miles D, Gianni L, Krop I, Welslau M, Baselga J & Blackwell K (2012) Trastuzumab emtansine for HER2-positive advanced breast cancer. The New England Journal of Medicine 367:1783–1791.Vilas-Bôas GT, Peruca AP, Arantes OM. (2007). Biology and taxonomy of Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis. Can. J. Microbiol. 53, 673-687.Xu C, Wang BC, Yu Z, Sun M (2014) Structural insights into Bacillus thuringiensis Cry, Cyt and parasporin toxins. Toxins. 6(9):2732-70.Yamashita S, Akao T, Mizuki E (2000) Characterization of the anti-cancer cell parasporal proteins of a Bacillus thuringiensis isolate. Can J Microbiol 46: 913-9Yamashita S, Katayama H, Saitoh H, Akao T, Park Y, Mizuki E, Ohba M Ito A (2005) Typical Three-Domain Cry Proteins of Bacillus thuringiensis Strain A1462 Exhibit Cytocidal Activity on Limited Human Cancer Cells. Journal of Biochemistry, 138, 663-672.Yasutake K, Dinh N, Kagoshima K, Uemori A, Ohgushi A, Maeda M, Mizuki E, Man Y, Ohba M (2006) Occurrence of parasporin-producing Bacillus thuringiensis in Vietnam. Can. J. Microbiol. 52: 365–372.Yasutake K, Uemori A, Kagoshima K, Ohba M (2007) Serological identification and insect toxicity of Bacillus thuringiensis isolated from the island Okinoerabu-jima, Japan. Appl. Entomol. Zool. 42 (2): 285–290Yasutake K, Uemori A, Binh N, Mizuki E, Ohba M (2008) Identification of Parasporin Genes in Vietnamese Isolates of Bacillus thuringiensis. Z. Naturforsch. 63: 139-143.PublicationORIGINALIdentificaciónyCaracterizacióndeParasporinas,obtenidasdeAisladosdeBacillussppNativosdeSuelosColombianos.pdfIdentificaciónyCaracterizacióndeParasporinas,obtenidasdeAisladosdeBacillussppNativosdeSuelosColombianos.pdfDocumento Principalapplication/pdf1937865https://repositorio.udes.edu.co/bitstreams/9692b120-3a8d-4d0e-9df6-8e6d786ecee4/downloadfb9bc65644b5e76bd994bad9ce5e9596MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-859https://repositorio.udes.edu.co/bitstreams/ecc4de6f-565b-4e72-b7d4-ace1aaa19cd1/download38d94cf55aa1bf2dac1a736ac45c881cMD52TEXTIdentificaciónyCaracterizacióndeParasporinas,obtenidasdeAisladosdeBacillussppNativosdeSuelosColombianos.pdf.txtIdentificaciónyCaracterizacióndeParasporinas,obtenidasdeAisladosdeBacillussppNativosdeSuelosColombianos.pdf.txtExtracted texttext/plain101665https://repositorio.udes.edu.co/bitstreams/cb417a64-8444-43d3-a737-0a0625ddd0e8/downloadc4e3eb32fea9b8227436b597a2240c8dMD53THUMBNAILIdentificaciónyCaracterizacióndeParasporinas,obtenidasdeAisladosdeBacillussppNativosdeSuelosColombianos.pdf.jpgIdentificaciónyCaracterizacióndeParasporinas,obtenidasdeAisladosdeBacillussppNativosdeSuelosColombianos.pdf.jpgGenerated Thumbnailimage/jpeg5676https://repositorio.udes.edu.co/bitstreams/ee92dcc3-7c7a-4b6c-a856-6658ba9c5e8d/downloadec5cd2a8ff30fc813f8e6a39a5d08679MD54001/5520oai:repositorio.udes.edu.co:001/55202022-10-25 09:55:28.746https://creativecommons.org/licenses/by-nc-nd/4.0/Derechos Reservados - Universidad de Santander, 2021https://repositorio.udes.edu.coRepositorio Universidad de Santandersoporte@metabiblioteca.comTGljZW5jaWEgZGUgUHVibGljYWNpw7NuIFVERVMKRGlyZWN0cmljZXMgZGUgVVNPIHkgQUNDRVNPCgo= |