摘要
[目的]构建可将甲状腺球蛋白抗原决定簇基因转染入真核细胞的重组载体。[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pcDNATM3．1D／V5-His- 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载体经鉴定准确无误。[结论]成功的构建了带有目的基因的真核表达质粒Tg-pcDNATM3．1D／V5-His- TOPO(?)。
[Objective] The recombined vector that can transfer thyroglobulin （Tg） epitopes gene into cells was construeted. [Methods] Total RNAs were extracted from human thyroid tissues 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 peDNA^TM 3.1D/VS-His-TOPO and then identified by PCR, restriction endonuclease and sequencing. [Resuits] The recombined vector with Tg epitopes gene was correct by identifying. [Conehusion] The eukaryotie expression vector Tg-pcDNA^TM3.1D/V5-His-TOPO with objective gene was constructed successfully.
出处
《武警医学院学报》
CAS
2006年第1期10-13,共4页
Acta Academiae Medicinae CPAPF
