T细胞淋巴瘤TCRγV_1重排基因真核表达载体的构建

Construction of T-cell Receptor γV_1 Rearrangement Gene Expression Vector in Eucaryotic Cells

目的 构建含TCRγV1重排基因的真核表达质粒。方法 用PCR法扩增含XbaI与BglII酶切位点的TCRγV1基因序列 ,对VR10 12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DH5α ,对重组质粒经序列测定 ,称VR10 12 /TCRγ。结果 已构建的质粒VR10 12 /TCRγ经序列测定含有完整的TCRγV1基因片段。结论 VR10 12 /TCRγ表达载体的构建为基因治疗淋巴瘤奠定了基础。

Objective This study was designed to construct the eucaryotic expression vector of TCRγV_1 rearrangement gene.Methods TCRγV_1 rearrangement gene containing Xba I and BglII endoenzyme sites was obtained using PCR ;double enzyme digestion was conducted for vector VR1012 and PCR product of TCRγV_1 gene; Both fragments were connected using ligase and transferred to E.coli-DH5α ; Reconstitute plasmid sequence was examined by auto-sequencing assay and was named as VR1012/TCRγ. Conclusion Reconstitution of VR1012/TCRγ vector lay a foundation for T-cell lymphoma gene therapy .