论文部分内容阅读
目的 研究我国G2和G3型轮状病毒主要中和抗原VP7基因在重组腺病毒中的表达。方法 在前期成功表达G1型VP7的基础上 ,选用我国G2和G3型主要流行株 97S4 3和 97S4 8VP7基因 ,用非复制型腺病毒载体对上述基因进行表达。结果 获得了表达我国G2型和G3型人轮状病毒流行株VP7基因的非复制型重组腺病毒rvAdG2VP7和rvAdG3VP7,应用PCR及Southernblot技术证实在重组腺病毒中整合有轮状病毒G2型VP7和G3型VP7基因 ,RT PCR证明重组腺病毒在感染的 2 93细胞内均能有效地转录插入基因 ,Westernblot检测到轮状病毒VP7基因的表达。结论 这一工作为进一步进行动物实验 ,发展多价轮状病毒疫苗打下了基础。
Objective To study the expression of major neutralizing antigen VP7 gene of G2 and G3 rotavirus in recombinant adenovirus. Methods Based on the successful expression of G1 type VP7 in the early stage, 97S4 3 and 97S4 8VP7 genes of major strains G2 and G3 in our country were selected and the non-replicating adenoviral vector was used to express these genes. Results The non-replicating recombinant adenovirus rvAdG2VP7 and rvAdG3VP7 expressing VP7 gene of human rotavirus type G2 and G3 were obtained. PCR and Southern blotting were used to confirm that the recombinant adenoviruses were integrated with rotavirus type G2 VP7 and G3 Type VP7 gene. RT-PCR proved that the recombinant adenovirus could efficiently transcribe the inserted gene in all 293 cells, and the expression of rotavirus VP7 gene was detected by Western blot. Conclusion This work laid the foundation for further animal experiments and development of multivalent rotavirus vaccine.