密码子优化提高重组腺病毒中人轮状病毒VP6基因表达量的研究

Improved expression of human rotavirus VP6 antigen in the recombinant adenoviruse by codon optimization

目的 通过密码子优化提高重组腺病毒中人轮状病毒VP6基因的表达量.方法 人工合成按照人的优势密码子优化的人轮状病毒VP6基因,包装重组腺病毒后,与未优化的相同基因的重组腺病毒用免疫荧光和Western Blot方法比较其表达量.结果 密码子优化的人轮状病毒VP6基因,与未优化的相同基因的重组腺病毒比较,其蛋白表达量大大提高.结论 密码子优化确实提高了重组腺病毒中人轮状病毒VP6基因的表达量,可望满足腺病毒载体疫苗的研发需要.

Objective To increase the recombinant adenovirus vector mediated human rotavirus VP6 gene expression through coden optimization.Methods We have artificially synthesized VP6 gene of group A human rotavirus according to the human biased codon.The modified gene was transfected into 293 cells using adenovirus vector and the gene product,the respective protein was produced.The expression level of optimized gene and wild type gene was detected by Immunofluorescence（IF） and Western Blot.Result A remarkable increase of the expression level of optimized VP6 gene in comparison with the wild-type control.Conclusion The coden optimization indeed help increasing the recombinant adenovirus mediated human rotavirus gene expression,which indicated the potential application of such recombinant adenoviruses in the development of adenoviral-vectored rotavirus vaccines.