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Biotechnol Appl Biochem ; 59(3): 245-53, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23586835

RESUMO

Nef is one of the accessory proteins of human immunodeficiency viruses. Here, we noted that the relative expression level of Nef(NL4-3) is much lower than that of NefJR-CSF in HEK293 cells. By evaluating the expression level using a Nef mutant, it was indicated that amino acids 129-206 of Nef(NL4-3), that is, the C-terminal region named NLAA129-206, could contain the region responsible for the induction of the low protein expression level. In addition, the expression levels of the enhanced green fluorescent protein and Renilla luciferase became extremely low with the fusion of NLAA129-206. Interestingly, the NLAA129-206-corresponding sequences of other Nef variants with relatively high expression levels also induced the extremely low protein expression level by fusion. These results suggest that the C-terminal region of Nef can generally induce an extremely low protein expression level. Here, we propose that the C-terminal region of Nef could become an excellent tool for the induction of an extremely low expression level of arbitrary proteins by attachment as fusion proteins.


Assuntos
Regulação Viral da Expressão Gênica/genética , Produtos do Gene nef/genética , HIV-1/genética , Proteínas Recombinantes de Fusão/genética , Western Blotting , Citometria de Fluxo , Produtos do Gene nef/metabolismo , Vetores Genéticos/genética , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Células HEK293 , Humanos , Luciferases de Renilla/metabolismo , Mutação , Plasmídeos/genética , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , Proteínas Recombinantes de Fusão/metabolismo
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