摘要
目的 克隆K ras癌基因 ,测定其序列及突变类型 ,构建逆转录病毒表达载体。方法 通过逆转录 聚合酶链式反应 (RT PCR)方法从胰腺癌细胞株中克隆K ras基因 ,与T Vector载体连接后测序。构建含K ras基因的逆转录病毒载体。结果 胰腺癌细胞株Bxpc 3、Sw1990未及点突变 ,Patu898812位密码子的GGT突变为GTT ,13、6 1位无突变。酶切证明逆转录病毒表达载体构建成功。结论 K ras癌基因的成功克隆及逆转录病毒表达载体的构建 ,为胰腺癌的免疫治疗研究奠定了基础。
Objective To clone K-ras oncogene,detective its mutation and construct retroviral vector. Methods There was special expression of K-ras oncogene point mutation in pancreatic cancer,K-ras oncogene was coloned from pancreatic cancer line Bxpc-3、Sw1990、Patu8988 by using RT-PCR method.By means of T-A extension cloning,the gene was ligated with T-Vector and sequenced.To construct retroviral vector constisting of K-ras oncogene. Results There were none of point mutation in pancreatic cancer line Bxpc-3、Sw1990 and 13th、61th point mutation in Patu 8988. We found GGT→GTT in 12th codon in Patu 8988 line. According to the double digestion,it was comfirmed that retroviral vector constisting of K-ras oncogene had been constructed successfully. Conclusion The successful clone of K-ras oncogene and construction of retroviral vector lays the foundation for the immunotherapy of pancreatic cancer.
出处
《苏州大学学报（医学版）》
CAS
2004年第2期144-146,共3页
Suzhou University Journal of Medical Science
