肺癌相关基因LSCC-3真核表达载体的构建及其表达

Construction and expression of eukaryotic vector carries total ORF of LSCC-3 in lung caner cell

目的构建含有肺癌相关基因LSCC-3全长ORF片段的真核表达载体,并验证其在细胞内的表达。方法PCR方法获取LSCC-3基因全长ORF片段,构建含有上述片段的真核表达载体(含3’端绿色荧光蛋白尾)并验证、纯化;将该真核表达载体转染人肺腺癌PAa细胞系,在倒置荧光显微镜下对LSCC-3基因的表达进行观察和验证。结果成功将LSCC-3基因全长ORF片段装入真核表达载体pLPS-3’EGFP(pLPS-LSCC-3-3’EGFP)并转染人肺腺癌PAa细胞系,在倒置荧光显微镜下观察到了核外细胞浆内LSCC-3基因蛋白表达产物的存在。结论成功构建了LSCC-3真核表达载体pLPS-LSCC-3-3’EGFP,该载体可在肺癌细胞胞浆内顺利表达。

Objective To construct a eukaryotic vector carries a segment of the total ORF of LSCC-3, then to investi-gate the expression of this vector.Methods Got the total ORF of gene LSCC-3 by a PCR method, Transferred this segment in-to a eukaryotic vector which has a 3＇-EGFP tail, Transfected lung cancer PAa cells by this eukaryotic vector and then investi-gated its intracellular expression under fluorescent microscopy.Results Total ORF of gene LSCC-3 was gotten and the eu-karyotic vector carries ORF of LSCC-3 was constructed successfully, And the expression of this vector was found in the cyto-plasm after the lung cancer PAa cells had been transfected.Conclusion pLPS-LSCC-3-3＇EGFP,an eukaryotic vector carries total ORF of LSCC-3 is constructed successfully and its intracellular expression in the cytoplasm is confirmed.