中国医学科学杂志(英文版)
中國醫學科學雜誌(英文版)
중국의학과학잡지(영문판)
CHINESE MEDICAL SCIENCES JOURNAL
2001年
3期
129-134
,共6页
孙茂盛%昝云红%马雁冰%张光明%杜秋江%戴长柏
孫茂盛%昝雲紅%馬雁冰%張光明%杜鞦江%戴長柏
손무성%잠운홍%마안빙%장광명%두추강%대장백
Objective. Using a recombinant human adenovirus to express modified VP4 gene of rotavirus SA11 strain.``Methods. A whole VP4 gene was obtained with PCR and induced the signal peptide at the gene N terninal.``The chimera gene was cloned into pCMV plasmid that consists of human cytomegalovirus promoter, and then the gene was cloned to the transfer vector of human adenovirus type 5. Homologous recombination was performed by co-transfection to 293 cell lines with recombinant plasmid and viral genome using CaPO4 precipitation.``Results. No mutation was found in the whole VP4 gene sequence of 2362 base pair. The expressed product in recombinant adenovirus was confirmed to be specific and more antigenicity by indirect immunofluorescence assay. Both the Western blot and immunoprecipitation assay showed that the molecular mass of the expressed protein was higher than the wild type VP4 protein, and that the modified product was corresponding to a glycosylation of VP4 protein.``Conclusion. To modify the target gene might be an effective method to enhance the stability, antigenicity and immunogenicity of expressed protein.``