莱茵衣藻叶绿体PsbA启动子的克隆及其活性验证

Amplification of PsbA Promoter Fragment from Chloroplast of Chlamydomonas Reinhardtii and Its Verification of Action

为验证莱茵衣藻叶绿体基因PsbA启动子活性,提取了莱茵衣藻总DNA。设计基因PsbA启动子引物,以总DNA为模板,利用PCR法扩增,然后与质粒P64D连接。将重组质粒转入大肠杆菌Dh5α。用氨苄青霉素和壮观霉素筛选,进行活性验证。结果成功地从莱茵衣藻基因组中克隆出PsbA启动子片段(1 161 bp),获得了氨苄青霉素和壮观霉素抗性菌落,表明该序列具有启动子活性。

To verify the activity of gene PsbA in chloroplast of Chlamydomonas reinhardtii,it is extracted the whole DNA of the Chlamydomonas reinhardtii and design the gene PsbA promoter primer.Take the DNA as a template,the fragment by PCR is amplified,then connect the fragment with plasmid p64D.The recombinant plasmid in E.coli Dh5α is leaded.The colony with ampicillin and spectinomycin is screen out.Then the activities are verified.Last the fragment（1 161 bp） is cloned successfully from chloroplast of Chlamydomonas reinhardtii.It is acquired the resistance colony of ampicillin and spectinomycin.It is testified that this sequence is of the promoter activities.