【摘要】 从人转移型胃癌裸鼠移植瘤中抽提出总RNA,分离出多聚腺苷酸RNA,合成双链cDNA,接上ECoRI接头,以pT7T3 18U质粒为载体,以大肠杆菌NM522为受体菌,构建了人转移型胃癌裸鼠移植瘤cDNA文库。该文库含1.5×10~6转化子,插入片段cDNA长度为0.4～6.0Kb,重组效率为80％,并用四种癌基因探针筛选了阳性克隆。更多还原

【Abstract】 Total RNA were extracted from the transplanted human primary gastric cancertissue in nude mice. The poly (A～+) RNA were isolated, dsDNA (0.4-6.0Kb) were syn-thesized and linked with ECoRI adaptor, pT7T318 U plasmid as vector and E Coli NM 522 as host cells were used to construct a cDNA library. The result showed thatthis cDNA library contained 1.5×10～6 transforments, screening with ～82P-α-tubulinprobe about 0.05% of the clones were positive. Among the positive clones about60% corresponded to the full length mRNA of α-tubulin. We have screened positiveclones of C-H-ras, N-ras, v-K-ras and C-myc in 4×10～5 transforments, the positiverate of C-H-ras and N-ras was much higher than the latter two.更多还原