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目的 建立转染野生型nm2 3 H1 基因的人大细胞肺癌细胞株L9981 nm2 3 H1 。方法 应用基因克隆技术构建逆转录病毒载体pLXSN nm 2 3 H1 EGFP。脂质体法转染PA3 17细胞 ,得到逆转录病毒 ,并将病毒感染L9981细胞 ,获得L9981 nm 2 3 H1 。应用PCR和Westernblot检测L9981 nm2 3 H1 细胞中nm2 3 H1基因及其蛋白表达。结果 成功构建了逆转录病毒载体pLXSN nm2 3 H1 EGFP。nm2 3 H1 cDNA导入到L9981细胞株中 ,并检测到L9981 nm 2 3 H1 细胞中有nm 2 3 H1 蛋白表达。结论 nm 2 3 H1 基因在细胞株L9981 nm2 3 H1 中能持续、稳定和高效表达
Objective To establish a human large cell lung cancer cell line L9981 nm23 H1 transfected with wild type nm2 3 H1 gene. Methods The retroviral vector pLXSN nm 2 3 H1 EGFP was constructed by gene cloning technique. Lipofectamine was transfected into PA3 17 cells to obtain retroviruses, which were then infected into L9981 cells to obtain L9981 nm 2 3 H1. The expression of nm23 H1 gene and its protein in L9981 nm23 H1 cells was detected by PCR and Western blot. Results The retroviral vector pLXSN nm2 3 H1 EGFP was successfully constructed. The nm23 H1 cDNA was introduced into L9981 cell line and the expression of nm23 H1 protein in L9981 nm23 H1 cells was detected. Conclusion The nm 2 3 H1 gene can be consistently, stably and efficiently expressed in L9981 nm2 3 H1 cell line