鼠源抗HFRSV衣壳蛋白McAbF3株单链抗体基因克隆构建及表达

为了获得原抗ＨＦＲＳＶ衣壳蛋白ＭｃＡｂＦ３＾１株轻链可变区基因，由连接肽体外连接获得单链抗体基因，在大肠杆菌中表达，从鼠源抗ＨＦＲＳＶ衣壳蛋白ＭｃＡｂＦ３株细胞中分离总ＲＮＡ，以ｏｌｉｇｏ（ｄＴ）１８为引物逆转录成ｃＤＮＡ，通过ＰＣＲ扩增出抗体的轻链（ＶＬ）和重链可变区（Ｖ１１）基因，由连接本外连接获得单链抗体（ＳｅＦｖ）基因。将此单链抗体（ＳｅＦｖ）基因插入原核表达载体ＰＥＴ２８ａ，经大肠杆菌（

Construction and expression of mouse anti-HFRSV capsid protein single-chain antibody

Constructing recombinant mouse anti hemorrhagic fever with renal virus syndrome(HFRS) single chain antibody gene,and expressing it in E.coli.The total RNA was separated,the cDNAs of variable heavy chain(V H) and variable light chain(V L) region were synthesized by PCR,the DNAs of heavy and light chain were jointed together via a linker to get mouse anti HFRSV ScFv gene,and then the ScFv gene was inserted into plasmid PET28a.The ScFv gene of mouse anti HFRSV is 741 bp,the obtained recombinant plasmid was transformed into E.coli BL21(DE3),and expressed by IPTG inducing.The expressed production was analyzed by SDS PAGE and Western blotting.Its molecular weight is 29 000 ,the expressed product could specifically react with rabbit anti mouse IgG.