丙肝病毒融合抗原基因导入烟草叶绿体及转化株同质化的研究

摘要

植物生物反应器的研究已成为当前基因工程领域的一个新生长点。本实验用PCR方法，从丙型肝炎病毒cDNA文库中克隆了非结构NS3区基因片段及核心抗原C区基因片段，并在两段基因间加入连接肽SPGS的密码子序列，构建成融合抗原基因NS3-C。将该融合基因转入烟草叶绿体转化及表达载体中，通过基因枪转化法得到转基因植株。对NS3-C融合基因转化植株进行PCR及Southern杂交试验分析，证明外源基因已整合到烟草叶绿体基因组中；对转化植株的Southern杂交试验还表明，通过不断地分化筛选培养，可以提高转化植株的同质化水平。%Spectinomycin-resistant calli and shoots were obtained from tobacco leaves bombarded with pSS3 DNA which contains the NS3-C chimeric antigen gene of hepatitis C virus. Further molecular screening on the transformed tobacco plants such as Southern blot and PCR amplifying were performed, which indicated that foreign gene had been integrated into the chloroplast genome. And Southern blot also showed that the homogenization degree will increase after repeatedly selected on spectinomycin medium. All of the above results showed that stable tobacco chloroplast genetic transformation system had been established, which provided a new way for plant engineering.