CAJ | 학술논문

目的对前期制备的以Ad41为载体的可表达A组轮状病毒VP6基因的非复制型重组腺病毒进行连续传代,观察其遗传稳定性,为进一步研发疫苗做准备.方法在293TE7细胞上,对本室前期构建的重组腺病毒rvAd41-VP6 (o)连续传代14代,每隔2代通过PCR鉴定VP6(o)基因的插入,通过Western Blot技术鉴定目的蛋白的表达.结果 PCR分析表明,rvAd41-VP6(o)在连续传代的过程中,其基因组中都有特异性的VP6基因稳定整合；Western Blot证实了rvAd41-VP6 (o)可稳定表达VP6,这说明rvAd41-VP6(o)有较好的遗传稳定性.结论重组腺病毒rvAd41-VP6 (o)在体外具有良好的生物学性质,可用于下一步动物免疫研究.
목적 대전기제비적이Ad41위재체적가표체A조륜상병독VP6기인적비복제형중조선병독진행련속전대,관찰기유전은정성,위진일보연발역묘주준비.방법 재293TE7세포상,대본실전기구건적중조선병독rvAd41-VP6 (o)련속전대14대,매격2대통과PCR감정VP6(o)기인적삽입,통과Western Blot기술감정목적단백적표체.결과 PCR분석표명,rvAd41-VP6(o)재련속전대적과정중,기기인조중도유특이성적VP6기인은정정합；Western Blot증실료rvAd41-VP6 (o)가은정표체VP6,저설명rvAd41-VP6(o)유교호적유전은정성.결론 중조선병독rvAd41-VP6 (o)재체외구유량호적생물학성질,가용우하일보동물면역연구.
Objective To investigate the genetic stability of non-replicating recombinant adenovirus which used Ad41 as vector and could express VP6 gene of group A rotavirus during continous passage,in order to develop the vaccine of rotavirus.Methods The recombinant adenovirus rvAd41-VP6 (o) was prepared by our laboratory early,it then was continuously propagated on 293TE7 cells for 14 passages.After that samples of the infected cells were collected at every 2 passages for the detection of the integration of the VP6 gene by PCR,and the expression of the target protein was detected by Western Blot analysis.Results Analysis by PCR revealed that,there was stable integration of specific VP6 gene in the rvAd41-VP6 (o),Western Blot analysis confirmed that rvAd41-VP6 (o) could stably expressed the group-specific antigen structural protein VP6 (o),and it had preferable genetic stability.Conclusion The recombinant adenovirus rvAd41-VP6(o) which could stably express the VP6(o) gene had favorable biological property in vitro,and it has provided a basis for further research of animal immunization.