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Using Bcl-xL anti-apoptotic protein for altering target cell apoptosis
Iz, Sultan Gülçe; Çalimlioglu, Beste; Gürhan, Saime Ismet Deliloglu.
Afiliação
  • Iz, Sultan Gülçe; Ege University. Engineering Faculty. Department of Bioengineering. Bornova. TR
  • Çalimlioglu, Beste; Ege University. Engineering Faculty. Department of Bioengineering. Bornova. TR
  • Gürhan, Saime Ismet Deliloglu; Ege University. Engineering Faculty. Department of Bioengineering. Bornova. TR
Electron. j. biotechnol ; 15(5): 2-2, Sept. 2012. ilus, tab
Artigo em Inglês | LILACS | ID: lil-657661
Biblioteca responsável: CL1.1
ABSTRACT

Background:

Altering target cell apoptosis is one of the challenging ideas of biotechnological applications. There are several applications of over expressing Bcl-xL anti-apoptotic protein from recombinant protein production to DNA vaccination strategies. The aim of the present study is to evaluate the anti-apoptotic efficacy of Bcl-xL expressing dual promoter plasmid system as a candidate to be used for recombinant protein production and DNA vaccination approaches. For this purpose, Bcl-xL anti-apoptotic protein gene was inserted in a dual expressing vector system in frame with EGFP (enhanced green fluorescence protein) after IRES (internal ribosomal site). The plasmid has a multiple cloning site after CMV (cytomegalovirus promoter) left empty to be inserted a biopharmaceutical protein gene region or DNA vaccine antigens.

Results:

In order to determine the anti-apoptotic efficacy of Bcl-xL inserted dual expressing vector, BHK-21 cells were transfected both with this plasmid and empty vector as control. Apoptosis was stimulated by several apoptosis inducing agents and serum deprivation in the transfected cells for 48 hrs. Cells expressing Bcl-xL protein in frame with EGFP were determined by flow cytometry as an indicator of cell viability. Additionally, apoptosis were determined by intracellular cleaved Casp 3 staining in Bcl-xL expressing EGFP positive cells. The dual expression plasmid bearing Bcl-xL anti-apoptotic protein prolonged the cell survival rate and protected cells from apoptosis upon apoptosis induction by doxorubicin and camptothecin in which the anti-apoptotic efficacies are inhibited through over expressing of Bcl-xL. pIRES2EGFP/Bcl-xL transfected cell ratio was significantly higher compared to empty vector transfected cells (P < 0.001). In contrast, apoptotic cell ratio was significantly lower in pIRES2EGFP/Bcl-xL transfected cell population compared to empty vector transfected cells (P < 0.001).

Conclusion:

In conclusion...
Assuntos


Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: LILACS Assunto principal: Apoptose / Proteína bcl-X Idioma: Inglês Revista: Electron. j. biotechnol Assunto da revista: Biotecnologia Ano de publicação: 2012 Tipo de documento: Artigo / Documento de projeto País de afiliação: Turquia Instituição/País de afiliação: Ege University/TR

Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: LILACS Assunto principal: Apoptose / Proteína bcl-X Idioma: Inglês Revista: Electron. j. biotechnol Assunto da revista: Biotecnologia Ano de publicação: 2012 Tipo de documento: Artigo / Documento de projeto País de afiliação: Turquia Instituição/País de afiliação: Ege University/TR
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