关键词: 人呼吸道合胞病毒, M2-1基因, 肺癌细胞, 脂质体, RT-PCR

Abstract: Objective Background and human respiratory syncytial virus(hRSV) is a most familiar pathogen for severe respiratory tract infection in infants and young children, and it is found that it is also an important pathogen in adults in recent years.In this study, we try to construct a eukaryotic cell expressing vector containing the M2-1 gene of the HRSVand then try to transfect the lung cancer cell lines PAa and A549, so that we can continue studying the relationship between HRSVand lung cancer.Methods To construct the vector pXJ-41/M2-1 with gene reconstructing technique.Then check it through DNA-sequencing, PCRand enzyme-cutting.To transfect this vector into lung cancer cell lines mediated by liposome LipofectamineTM 2000 and screen out high expressing cell strains by RT-PCRin reference to the β-actin, which is an endogenous reference gene.Results 650 bp segment was obtained by PCRwith the recombinant as a template or by digesting the recombinant with EcoRIand XholI.The DNAsequencing result indicated that the M2-1 gene is high-conservative.Cell strains of PAa and A549 which can stably express M2-1 protein were acquired.Conclusion The eukaryotic cell expressing vector pXJ-41/M2-1 was successfully constructed.The acquirement of PAa and A549 cell strains expressing M2-1 protein would make it possible to study the influence of hRSVon lung cancer cells.

Key words: human respiratory syncytial virus, M2-1 gene, lung cancer cell, liposome, RT-PCR