构建N ras基因第 6 1位密码子突变基因 (N ras/ 6 1)的重组痘苗病毒 ,通过痘苗病毒载体所产生的蛋白的强烈免疫原性来提高变异N ras蛋白的肿瘤抗原性。方法 :利用双PCR方法得到第 6 1位突变的N ras/ 6 1基因片段 (5 0 0bp) ,将该基因片段插入痘苗病毒表达载体P110 8,通过CellFECTIN转染CV 1细胞 ,以PCR ,Westernblot方法鉴定重组痘苗病毒。结果 :通过tk-筛选、PCR鉴定 ,得了第 6 1位突变N ras/ 6 1基因的重组痘苗病毒rV N ras/ 6 1,Westernblot检测其在CV 1细胞中有表达。结论 :目前对突变ras基因的研究多着重于机理方面 ,本研究对研制有效的突变ras基因疫苗是一个有益的尝试 ,其免疫治疗作用有待于进一步研究。

To construct the recombinant vaccinia virus of mutant N ras/61 gene and enhance the immunogenecity of mutant N ras/61 protein produced by the recombinant vaccinia virus. Methods: N ras/61 gene was inserted into P1108 and transfected into CV 1 cell infected with vaccinia virus by Cell FECTIN. PCR and Western blot were used to identify the recombinants. Results: We get recombinant vaccinia virus rV N ras/61 by PCR and tk - selecting. The expression of N ras gene was detected by Western blot. Conclusion: This study is a test for studing effective vaccine of mutant N ras/61 gene.The efficacy in vivo of the N ras/61 vaccine in immunotherapy is under investgation.

本课题受山东省科委科研基金(951166307)资助