论文部分内容阅读
目的克隆肾癌G250抗原蛋白多糖(PG)区cDNA,为制备G250杂交瘤及抗G250的单克隆抗体奠定基础。方法从肾癌细胞786-0中提取总RNA,以RT-PCR方法扩增出全长360 bp的G250抗原PG区cDNA,将其与表达载体pCA13连接,转化大肠杆菌JM109,构建G250抗原PG区cDNA克隆。结果酶切及序列分析表明,与GenBank报道的G250抗原PG区cDNA序列完全一致。结论G250抗原PG区cDNA已成功地得到克隆。
OBJECTIVE: To clone the G2 region of rat kidney cancer proteoglycan (PG) cDNA and lay the foundation for preparation of G250 hybridoma and monoclonal antibody against G250. Methods Total RNA was extracted from human renal cell carcinoma cell line 786-0. The full length 360 bp cDNA of G250 antigen was amplified by RT-PCR and cloned into expression vector pCA13. The recombinant plasmid was transformed into E. coli JM109 to construct G2 region antigen region cDNA clone. Results The results of enzyme digestion and sequence analysis showed that the cDNA sequence of PG region of G250 antigen reported by GenBank was completely identical. Conclusion The G2 region antigen of PG has been successfully cloned.