摘要
以痘苗病毒HA基因为侧翼,将HIV-2gag基因部分序列(简称G1)和全部序列(简称G2)分别插入牛痘病毒A型包涵体(ATI)和串联10个(与HA基因反向)或16个(与HA基因同向)痘苗病毒P7.5组成的复合型启动子下游,构建了4个重组痘苗病毒表达载体质粒。经脂质体转染、鸡红细胞吸附试验筛选和免疫荧光鉴定,获得4株能稳定表达目的蛋白的重组痘苗病毒。实验结果表明,以HA基因为侧翼构建重组病毒的重组率约为0.1%,阳性重组率为25%~100%。实验中还发现,以HA基因为侧翼的重组痘苗病毒能够引起细胞融合,其可作为筛选重组痘苗病毒的另一种标志。Westernblot结果表明,表达的Gag蛋白能被HIV-2血清中的特异性抗体所识别,并能诱导小鼠产生抗HIV-2Gag抗体。
Four eucaryotic expression plasmids containing cDNA of HIV2 gag gene under the control of different hybrid promoters of pox virus and 10 or 16 tandemly repeated mutant early P7.5 were constructed, and transfected into BHK21 cells which were infected by wild type vaccinia virus. The recombinant rate of 0.1%, positive recombinant rate of 25%100% and the syncytium formation to great extent with flank sequence of HA gene of vaccinia virus were observed. Indirect immunofluorescence assay (IFA) and Western blot showed that all the recombinant vaccinia viruses expressed the Gag protein which can specifically response to HIV2 positive serum, and induce HIV2 Gag specific antibody in mice.
出处
《中国兽医学报》
CAS
CSCD
北大核心
1997年第6期570-574,共5页
Chinese Journal of Veterinary Science