摘要
利用RT-PCR技术获得登革2型病毒中国分离株(D_2-43)的prM基因片段。扩增产物经酚:氯仿抽提纯化之后,直接插入pT_7BlueT载体中,经DNA序列分析证明了所扩增片段序列的正确性。构建了痘苗病毒载体PJSA1175/prM,外源基因受痘苗病毒启动子P_(7.5K)的调控。脂质转染(Lipofection)法获得D_2-43株prM蛋白的重组病毒。荧光检测显示,重组痘苗病毒表达的prM蛋白分布于细胞表面。间接ELISA测定其滴度为1:4。
The prM gent fragment of Dengue 2 virus China isolate (D2-43) was obtained by RT-PCR technique. After extrated and purified with phenol: chloroform, the amplified product was directly inserted into pT7-Blue T vector. Nucleotide sequence of the amplified gene fragment was confirmed by DNA sequencing analysis. Then we constructed vaccinia virus recombinant plasmid JSA1175/prM in which the foreign geng was controlled by vaccinia virus promotor P7 5k. Using lipofectin-transfection method, we aquired D2-43 prM recombinant vaccinia virus. Indirect immunosorbent assay showed that the recombinant vaccinia virus expressed prM protein was anchored on cell surface. Result of enzyme-linked immunosorbent test showed that the antigen liter was 1 : 4.
出处
《生物技术通讯》
CAS
1998年第1期6-9,共4页
Letters in Biotechnology
