Effect of Codon Bias on the Gene Expression Efficiency in Vaccinia Virus Vector
To determine the relationship between codon bias and the gene expression efficiency in vaccinia virus vector, green fluorescent protein gene was recoded and synthesized according to the preferred codons by human and vaccinia virus, respectively. Then each of those synthesized genes was used to construct recombinant vaccinia virus (rVV). Immunofluorescence, Western blot and flow cytometry analyses were used to analyze the expression levels of synthetic genes in rVV-infected HeLa cells. It was also analyzed on the expression levels of the three green fluorescent protein genes with different codon bias in traditional plasmid vector pcDNA3 (transcribing target gene in nucleus) and SFV-based plasmid vector pSCA (transcribing target gene in cytoplasm). The results showed that all of these three target genes had expressed efficiently in vaccinia virus vector and the codon bias in those target genes just had slight impact on the expression efficiency. However, the G + C rich green fluorescent protein genes in pcDNA3 plasmid vector, including mGFP gene (G + C content is 51.5 % ) and EGFP gene (G+ C content is 61.8%) were expressed efficiently, but the A + T rich green fluorescent protein gene VGFP (A + T content is 73.1% ) exhibited very poor expression. In construct, all green fluorescent protein genes in pSCA plasmid vector had efficient and comparable expression in suitable host cells. These results demonstrated that vaccinia virus vector has a broad compatibility to the codon usage of target genes. The mechanism of this broad compatibility is mainly related to the cytoplasmic transcription of vaccinia virus. Our study provides very useful information on choosing vaccinia virus vector and other common eukaryotic vectors.
NETL
NSTL
万方数据
维普
