摘要
目的 获得稳定表达人呼吸道合胞病毒(h RSV) M2 - 1基因的人肺腺癌细胞系。方法 通过基因重组法构建h RSV M2 - 1基因真核表达载体,用脂质体将其转染人肺腺癌PAa和A5 4 9细胞,经G4 18筛选获得阳性表达细胞株,并用逆转录-聚合酶链反应(RT- PCR)和Western Blot进行验证。结果 得到了约6 5 0 bp的基因插入片段,DNA测序表明该基因高度保守。筛选出了稳定高量表达M2 - 1基因的PAa和A5 4 9细胞,并被证实有M2 - 1蛋白表达。结论 获得了稳定表达h RSV M2 - 1蛋白的PAa和A5 4
Objective To acquire lung adenocarcinoma cell lines which steadily expressing human respir atory syncytial virus(hRSV) M2-1 gene.Methods The vector M2-1 was constructed with gene reco nstructing technique.Then it was checkecl through DNA-sequencing,polymerase c hain reaction (PCR) and enzyme-cutting.This vector was transfected into lung a denocarcinoma cell lines PAa and A549 mediated by liposome Lipofectamine TM 2000 and screen out high expressing cell strains by reverse transcription-PCR( RT-PCR) and Western Blot.Results About 650bp segment was obtained.The M2-1 gene was high-conservative.The cell strains of P and A549 which stably expressing M2-1 gene were screened.Conclusion The P and A549 cell strains that expressing hRSV M2-1 protein are acquired.
出处
《山东医药》
CAS
北大核心
2005年第12期15-16,共2页
Shandong Medical Journal
基金
全军医药卫生科研基金课题 (No.0 1MA0 14 )
