摘要
目的:扩增甲状腺球蛋白抗原决定簇基因,构建可转化入大肠杆菌的重组表达载体。方法:用TRIzol一步法从甲状腺组织中提取总RNA,RT-PCR逆转录出cDNA,PCR扩增出目的基因,与pET102/D-TOPO载体进行拓扑克隆,对重组质粒进行PCR、酶切及测序鉴定。结果:甲状腺球蛋白抗原决定簇基因及其重组表达载体经鉴定准确无误。结论:成功构建了带有目的基因的原核表达载体。
Objective： The recombined the vector that can transfer thyroglobulin（Tg） epitopes gene into E.coli was constructed. Methods： Total RNAs were extracted from the human thyroid tissue of patient with Graves＇ disease（GD） by TRIzol reagent. The Tg epitopes gene was amplified by RT-PCR. The target gene was inserted into the expressive vectors of pET102/D-TOPO and then identified by PCR, restriction endonuclease and sequencing. Results： The recombined vector with Tg epitopes gene was correct by identification . Conclusion： The prokaryotic expression vector with target gene was constructed successfully.
出处
《天津医科大学学报》
2005年第4期550-553,共4页
Journal of Tianjin Medical University
