摘要
目的 为乙型肝炎病毒基因工程双特性抗体的研制奠定基础。方法 提取抗乙型肝炎病毒杂交瘤细胞总RNA ,利用鼠源性抗体可变区通用引物 ,经逆转录PCR扩增轻链可变区和重链可变区基因 ,用连接肽 (Gly4Ser) 3连接轻、重链可变区基因 ,构建单链抗体基因 ,并克隆pGEM -TEasy质粒 ,进行核苷酸序列分析。结果 分别得到了 4 14bp的轻链可变区基因片段和 3 93bp的重链可变区基因片段 ,序列分析表明轻链可变区属于小鼠kappa链第XX家族 ,重链可变区属于小鼠Ig链第IX家族。结论 克隆到序列正确的抗乙型肝炎病毒表面抗原单链抗体基因。
Objective To provide foundation for studying bispecific antibodies against HBV by genetic engineering.Methods Purifying the total RNA of the hybridomas against HBsAg, amplifying the variable region of light chain (VL) and variable region of heavy chain(VH) by RT- PCR through the universal primers of the variable region of mouse immunoglobulin,ligating VL and VH fragments by a peptide linker(Gly\-4Ser)\-3,constructing ScFv into pGEM-T Easy vector and analyzing its sequence by automatic sequencing.Results 414bp VL amplying fragment and 393bp VH product were obtained by RT-PCR, and 414bp of VL belonged to the family XX of K chain,393bp of VH belonged to the family IX of heavy chain.Conclusion The sequence of ScFv against HBsAg was successfully cloned.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2001年第9期773-774,共2页
Chinese Journal of Public Health
