PCR检定OSMcDNA转染细胞中基因组整合与转录

Identification of Human OSM cDNA Integrated into Genome of Mouse Melanoma Cells and Its mRNA Transcripts by PCR Methods.

用ＰＣＲ和ＲＴＰＣＲ方法对人ＯＳＭｃＤＮＡ转染的小鼠黑色素瘤细胞进行基因组整合和ｍＲＮＡ 转录的检定．基因组整合检定时， 采用与调控序列和ｃＤＮＡ 序列相对应的上、下游引物， 以连续的转录单位进行扩增， 能够更准确地反映整合与表达的关系； ｍ ＲＮＡ 检定时， 采用与ｃＤＮＡ序列和质粒克隆位点与加ｐｏｌｙＡ 信号之间序列相对应的上、下游引物，

Genomic integration and mRNA transcripts of human OSM cDNA in transfected mouse melanoma cells were identified by PCR and RT PCR methods.A sense primer for the regulatory sequence in carrier vector paired with an antisense primer for cDNA was used in integration analysis, a continuous transcription unit was amplified with the expected size in OSM cDNA transfected cells but not in the wild type or vector control cells, reflecting a more accurate relationship between integration and expression. In transcription analysis, a sense primer for cDNA paired with an antisense primer for a sequence between the multiple cloning site and polyA signal in carrier vector was used to distinguish exogenous transcripts from endogenous gene products . This method is convenient and specific in determining exogenous gene integration and expression in transfectants.