论文部分内容阅读
目的 :在本室前期工作的基础上构建汉滩病毒M基因G2片段与S基因 0 .7kb片段嵌合基因的重组腺病毒 .方法 :构建含有汉滩病毒G2S0 .7嵌合基因的转移载体pShuttle G2S0 .7,然后通过特异性的酶切将嵌合基因与腺病毒DNA相连 ,电转化E .coliJM1 0 9并用PCR方法进行筛选和鉴定 ,获得重组腺病毒Adeno G2S0 .7DNA ,转染HEK2 93细胞得到重组腺病毒 .进一步对重组腺病毒滴度和表达产物进行鉴定 .结果 :构建了含G2S0 .7嵌合基因重组腺病毒 ,滴度可达 1 0 1 3 ～ 1 0 1 5pfu/L ;该重组腺病毒感染HEK2 93细胞后 ,表达出可被抗汉滩病毒核蛋白及糖蛋白G2的特异性单抗 (mAb)所识别的融合蛋白 .结论 :利用腺病毒表达系统 ,成功地表达同时具有核蛋白及糖蛋白G2生物学活性的融合蛋白G2S0 .7,为进一步研究其免疫学特性奠定了基础
OBJECTIVE: To construct a recombinant adenovirus carrying the chimeric gene of Hantaan virus M gene and S gene of 0. 7kb based on the previous work in our hospital.Methods: The transfer vector pShuttle containing Hantaan virus G2S0.7 chimeric gene was constructed. G2S0.7, and then by specific digestion of the chimeric gene and adenovirus DNA, E. coliJM1 0 9 electroporation and screening and identification by PCR method to obtain recombinant adenovirus Adeno G2S0 .7 DNA transfected HEK2 93 cells The recombinant adenovirus was obtained, and the recombinant adenovirus titer and expression product were identified.Results: The recombinant adenovirus containing G2S0.7 chimeric gene was constructed with the titer of 1 0 1 3 ~ 1 0 1 5pfu / L, After HEK2 93 cells were infected with the recombinant adenovirus, the fusion protein recognized by mAb of anti-Hantaan virus nucleoprotein and glycoprotein G2 was expressed.Conclusion: The adenovirus expression system can successfully express HEK2 93 The fusion protein G2S0.7, which is the biological activity of nucleoprotein and glycoprotein G2, lays the foundation for further study of its immunological properties