鼠bcl-X_L基因在CHO细胞中的表达

Expression of murine bcl-X_L gene in CHO cells

目的 :构建携带鼠bcl XL 基因的真核表达载体 ,并将其在CHO细胞中表达 .方法 :用反转录PCR获取鼠bcl XL全长cDNA序列 ,将其克隆进pEGFP重组融合表达质粒 .脂质体法转染CHO细胞 ,荧光检测目的蛋白表达 .结果 :成功获得了鼠bcl XLcDNA ,构建了编码鼠bcl XL 的真核融合表达载体pEGFP bcl XL,转染CHO细胞后观察到融合蛋白表达 .结论 :鼠bcl XL 基因的克隆及融合表达 ,为进一步在高密度、大规模细胞培养中利用bcl

AIM: To construct eukaryotic expression vector containing murine bcl X L and EGFP fusion gene and to express it in CHO cells. METHODS: RT PCR was used to obtain murine bcl X L cDNA. Then the recombinant expression vector pEGFP bcl X L was constructed by cloning bcl X L cDNA into eukaryotic expression vector pEGFP. After transfection into CHO cells through lipofectamine 2000 TM , the expression of fusion gene was detected by fluoroscopy. RESULTS: The obtained murine bcl X L cDNA was consistent with that in GenBank. The eukaryotic expression vector pEGFP bcl X L was successfully constructed and expressed in CHO cells. CONCLUSION: The cloning and expression of murine bcl X L gene are of significance in further use of bcl X L gene in high density large scale cell culture.