Detecção, clonagem e análise in silico do gene parasporina 4 em linhagens de Bacillus thuringiensis
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Tipo de documento: | Trabalho de conclusão de curso |
Idioma: | por |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://hdl.handle.net/11449/214266 |
Resumo: | Bacillus thuringiensis (Bt) is a gram-positive bacterium that has its strains studied mainly for the control of insect pests, due to the insecticidal capacity of most of its proteins (Cry and Vip). However, recent studies indicate the presence of other proteins with little known insecticidal action, but of great importance in research involving this microorganism. These proteins called Parasporins (PS) have cytotoxic activity and have six subfamilies, being PS1, PS2, PS3, PS4, PS5 and PS6, considered potential drugs in the treatment of cancer. Given the high incidence of cancer cases in the world and the need for new treatment alternatives, this study aimed to characterize B. thuringiensis strains, belonging to the collection of the Laboratório de Genética de Bactérias e Biotecnologia Aplicada at FCAV - UNESP, as potential sources of the PS4 protein, being the least studied as a subfamily. The selection of strains was performed using the PCR (Polymerase Chain Reaction) technique with specific primers for the ps4 gene, followed by cloning and sequencing of the amplicons of the positive responses, in order to establish a sequence analysis in silico. compared to existing locations in the GenBank database. The present study appears to be the selection of a strain carrying the ps4 gene, called B. thuringiensis coreanensis, whose partial sequence of the same was carried out for identity analysis with the database sequences and the observation that the ps4 gene is located on a plasmid. Therefore, this strain can be a source of the gene for future expression experiments and characterization of the cytotoxic effects of Parasporin 4 on different types of tumors. |
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Detecção, clonagem e análise in silico do gene parasporina 4 em linhagens de Bacillus thuringiensisDetection, cloning and in silico analysis of the parasporin 4 gene in Bacillus thuringiensis strainsBacillus thuringiensisParasporinaCâncerCitotoxicidadeGenética molecularBacillus thuringiensis (Bt) is a gram-positive bacterium that has its strains studied mainly for the control of insect pests, due to the insecticidal capacity of most of its proteins (Cry and Vip). However, recent studies indicate the presence of other proteins with little known insecticidal action, but of great importance in research involving this microorganism. These proteins called Parasporins (PS) have cytotoxic activity and have six subfamilies, being PS1, PS2, PS3, PS4, PS5 and PS6, considered potential drugs in the treatment of cancer. Given the high incidence of cancer cases in the world and the need for new treatment alternatives, this study aimed to characterize B. thuringiensis strains, belonging to the collection of the Laboratório de Genética de Bactérias e Biotecnologia Aplicada at FCAV - UNESP, as potential sources of the PS4 protein, being the least studied as a subfamily. The selection of strains was performed using the PCR (Polymerase Chain Reaction) technique with specific primers for the ps4 gene, followed by cloning and sequencing of the amplicons of the positive responses, in order to establish a sequence analysis in silico. compared to existing locations in the GenBank database. The present study appears to be the selection of a strain carrying the ps4 gene, called B. thuringiensis coreanensis, whose partial sequence of the same was carried out for identity analysis with the database sequences and the observation that the ps4 gene is located on a plasmid. Therefore, this strain can be a source of the gene for future expression experiments and characterization of the cytotoxic effects of Parasporin 4 on different types of tumors.Bacillus thuringiensis (Bt) é uma bactéria gram-positiva que possui suas linhagens estudadas principalmente para o controle de insetos-praga, devido à capacidade inseticida da maioria de suas proteínas (Cry e Vip). Porém, recentes estudos apontam a presença de outras proteínas com ação inseticida pouco conhecida, mas de grande importância nas pesquisas envolvendo este microrganismo. Essas proteínas denominadas Parasporinas (PS) possuem atividade citotóxica e apresentam seis subfamílias, sendo PS1, PS2, PS3, PS4, PS5 e PS6, consideradas potenciais fármacos no tratamento do câncer. Dada a grande incidência de casos de câncer no mundo e a necessidade de novas alternativas de tratamento, o presente trabalho teve por objetivo a caracterização de linhagens de B. thuringiensis, pertencentes à coleção do Laboratório de Genética de Bactérias e Biotecnologia Aplicada da FCAV – UNESP, como potenciais fontes da proteína PS4, sendo essa a menos estudada dentre as subfamílias supramencionadas. A seleção das linhagens foi realizada por meio da técnica de PCR (Reação da Polimerase em Cadeia) com primers específicos para o gene ps4, seguida pela clonagem e sequenciamento dos amplicons das amostras positivas, no sentido de se estabelecer uma análise in silico das sequências obtidas em comparação àquelas existentes no banco de dados GenBank. O presente estudo permitiu a seleção de uma linhagem portadora do gene ps4, denominada B. thuringiensis coreanensis, cuja sequência parcial do mesmo permitiu a análise de identidade com as sequências do banco de dados e a observação de que o gene ps4 encontra-se localizado em um plasmídeo. Portanto, esta linhagem poderá ser fonte do gene para futuros experimentos de expressão e caracterização dos efeitos citotóxicos da Parasporina 4 frente a diferentes tipos de tumores.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)2019/21621-0Universidade Estadual Paulista (Unesp)Desidério, Janete Apparecida [UNESP]Universidade Estadual Paulista (Unesp)Santos, Thais Nayara Ferreira dos2021-08-31T18:41:50Z2021-08-31T18:41:50Z2021-07-22info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/bachelorThesisapplication/pdfhttp://hdl.handle.net/11449/214266porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESP2023-12-16T06:22:11Zoai:repositorio.unesp.br:11449/214266Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T20:30:22.856150Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Detecção, clonagem e análise in silico do gene parasporina 4 em linhagens de Bacillus thuringiensis Detection, cloning and in silico analysis of the parasporin 4 gene in Bacillus thuringiensis strains |
title |
Detecção, clonagem e análise in silico do gene parasporina 4 em linhagens de Bacillus thuringiensis |
spellingShingle |
Detecção, clonagem e análise in silico do gene parasporina 4 em linhagens de Bacillus thuringiensis Santos, Thais Nayara Ferreira dos Bacillus thuringiensis Parasporina Câncer Citotoxicidade Genética molecular |
title_short |
Detecção, clonagem e análise in silico do gene parasporina 4 em linhagens de Bacillus thuringiensis |
title_full |
Detecção, clonagem e análise in silico do gene parasporina 4 em linhagens de Bacillus thuringiensis |
title_fullStr |
Detecção, clonagem e análise in silico do gene parasporina 4 em linhagens de Bacillus thuringiensis |
title_full_unstemmed |
Detecção, clonagem e análise in silico do gene parasporina 4 em linhagens de Bacillus thuringiensis |
title_sort |
Detecção, clonagem e análise in silico do gene parasporina 4 em linhagens de Bacillus thuringiensis |
author |
Santos, Thais Nayara Ferreira dos |
author_facet |
Santos, Thais Nayara Ferreira dos |
author_role |
author |
dc.contributor.none.fl_str_mv |
Desidério, Janete Apparecida [UNESP] Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Santos, Thais Nayara Ferreira dos |
dc.subject.por.fl_str_mv |
Bacillus thuringiensis Parasporina Câncer Citotoxicidade Genética molecular |
topic |
Bacillus thuringiensis Parasporina Câncer Citotoxicidade Genética molecular |
description |
Bacillus thuringiensis (Bt) is a gram-positive bacterium that has its strains studied mainly for the control of insect pests, due to the insecticidal capacity of most of its proteins (Cry and Vip). However, recent studies indicate the presence of other proteins with little known insecticidal action, but of great importance in research involving this microorganism. These proteins called Parasporins (PS) have cytotoxic activity and have six subfamilies, being PS1, PS2, PS3, PS4, PS5 and PS6, considered potential drugs in the treatment of cancer. Given the high incidence of cancer cases in the world and the need for new treatment alternatives, this study aimed to characterize B. thuringiensis strains, belonging to the collection of the Laboratório de Genética de Bactérias e Biotecnologia Aplicada at FCAV - UNESP, as potential sources of the PS4 protein, being the least studied as a subfamily. The selection of strains was performed using the PCR (Polymerase Chain Reaction) technique with specific primers for the ps4 gene, followed by cloning and sequencing of the amplicons of the positive responses, in order to establish a sequence analysis in silico. compared to existing locations in the GenBank database. The present study appears to be the selection of a strain carrying the ps4 gene, called B. thuringiensis coreanensis, whose partial sequence of the same was carried out for identity analysis with the database sequences and the observation that the ps4 gene is located on a plasmid. Therefore, this strain can be a source of the gene for future expression experiments and characterization of the cytotoxic effects of Parasporin 4 on different types of tumors. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-08-31T18:41:50Z 2021-08-31T18:41:50Z 2021-07-22 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/bachelorThesis |
format |
bachelorThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/11449/214266 |
url |
http://hdl.handle.net/11449/214266 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
publisher.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
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Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
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