Improvement of human interferon HUIFNalpha2 and HCV core protein expression levels in Escherichia coli but not of HUIFNalpha8 by using the tRNA(AGA/AGG).
High-level expression from one particular heterologous gene in Escherichia coli generally requires the optimization of codon usage. Genes encoding for Hepatitis C virus core protein (HCcAg), human interferon alpha2 and 8 subtypes (HUIFNalpha2 and HUIFNalpha8) show a high content of AGA/AGG codons. These are encoded by the product of the dnaY gene in E. coli. The proteins used in this work have a high therapeutic value and were used as models for studying the effects of these rare codons on the efficiency of heterologous gene expression in E. coli. Expression plasmids were constructed to express any of these proteins and the dnaY gene product simultaneously in E. coli. After dnaY gene expression, HCcAg, and HUIFNalpha2 expression levels increased 5 and 3 times, respectively. However, HUIFNalpha8 expression was barely detected either supplying or not the additional dnaY gene product. These results suggest that the high frequency of AGA/AGG codons present in the HCcAg and HUIFNalpha2 genes could be one of the factors limiting its expression in E. coli. Nevertheless, for HUIFNalpha8 it seems that other factors prevail upon the lack of dnaY product. Data presented here for HCcAg and HUIFNalpha2 expressions proved the value of this approach to obtain therapeutic proteins in E. coli.
Division of Vaccines, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, C.P. 10600, Havana, Cuba.
Nelson
N
Acosta-Rivero
Julio C
JC
Sánchez
Juan
J
Morales
D000998:Antiviral Agents / Q000378:metabolism*; D001483:Base Sequence; D003062:Codon; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D016898:Interferon-alpha / Q000096:biosynthesis / Q000235:genetics*; D007372:Interferons / Q000096:biosynthesis / Q000235:genetics*; D010957:Plasmids; D012343:RNA, Transfer / Q000235:genetics; D014169:Transformation, Bacterial; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000235:genetics*
D000998:Antiviral Agents; D003062:Codon; C554164:IFNA8 protein, human; D016898:Interferon-alpha; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus; D007372:Interferons; D012343:RNA, Transfer