【摘要】 目的:构建含TCRγV1重排基因的真核表达质粒。方法:从人淋巴瘤Jurkat细胞中提取mRNA,用RT-PCR法扩增含BamH与Hind酶切位点TCRγV1基因序列,对pcDNA3载体及TCRγV1基因PCR产物经双酶切,用连接酶将两者连接并转化到大肠杆菌DH5a,对重组质粒经序列测定,称pcDNA3/TCRγV1。结果:电泳获得333bp的TCRγV1预期条带。结论:该条带测序证实为TCRγV1基因序列。更多还原

【Abstract】 Objective:To construct the eucaryotic exp rc ssion vector of TCRγV₁ rearrangement gene.Methods:The mRNA o f TCRγV₁ was extracted from human Jurkat cells:TCRγV₁ rearrangement gene c ontaining Bam HⅠ and HindⅢ endoenzyme sites was obtained using RT-PCR method. The expressing plsasmid pcDNA3 and TCRγV₁ gene were cleaved with 2 restrictio n endonuclease BaamHⅠ and HindⅢ;Both fragments were connected using ligase and transformed into bacteria DH5a;Clonies were screened and the recombinant was se quenced for identification.Result:The expected 333bp band of TCR γV₁ was obtained by eletrophoresis.Conclusion:The band is con firmed TCR V₁ gene array byalignment determination.更多还原