人重组pcDNA3人钠/碘同向转运体核表达质粒的构建

Construction of recombinant human eukaryotic expression vector pcDNA3-hNIS

目的:获取、构建并鉴定携带人钠/碘同向转运体(hNIS)基因的pcDNA3.0质粒载体。方法:利用逆转录和聚合酶链反应获取Graves甲亢患者甲状腺NIS基因可编码区片段。与pUCm-T质粒载体连接、经转化、蓝白斑筛选、酶切鉴定,序列测定鉴定,获得pUCm-T-重组质粒。pUCm-T-重组质粒亚克隆得到pcDNA3-hNIS重组质粒。结果:所构建的pcDNA3-hNIS重组质粒中的hNIS基因序列与文献报道序列一致。结论:成功构建pcDNA3-hNIS重组质粒。

Objective： To construct and identify recombinant human eukaryotic expression vector with the Sodium/ Iodide symporter （ NIS ） genes. Methods ： Human Sodium/ Iodide symporter genes cDNA were amplified from thyroid tissue of the patient suffering from Graves disease by Rt - PCR , arid cloned into pUCm - T - hNIS , blue/ white color selection and restriction enzyme analysis, pUCm- T- hNIS was sequenced and subcloned into pcDNA3 -hNIS. Results：The results showed that the sequence data of hNIS gene in recombinant plasmid corresponded to those of reference report. Conclusion ：Successfully constructed pcDNA3 - hNIS recombinant plasmid can provide academic evidence for further research.