摘要
目的 研究肾癌中的信号传导尤其是Wnt/Frizzled信号通路在肾癌发生、发展中的作用 ,寻找具有生物意义的小肽 ,构建肾癌的酵母双杂交文库。 方法 提取肾癌组织的mRNA ,逆转录合成cDNA ,SalI/NotI酶切后基因重组法定向克隆于穿梭质粒pPC86中 ,转化DH5α菌后扩增。 结果 文库大小为 6× 10 6,SalI/NotI酶切鉴定 ,插入cDNA片段多为 1～ 3kb。 结论 肾癌pPC86酵母双杂交文库的构建成功为研究肾癌发生机理、诊断和治疗 ,尤其是信号分子通路治疗提供了有效的工具。
Objective To investigate cell signal transduction in renal carcinoma especially the Wnt/Frizzled pathway. Methods mRNA of renal neoplasm was isolated and cDNA sythsized by reverse transcription.After digested by Sal I and Not I ,ds cDNA was subcloned into the transfer plasmid pPC 86.Finaly,the renal cancer yeast two library has been constructed. Results The transformation rate of the library is 6×10 6.12 positive clones were chosed and minicrepted.Digested by Sal I and Not I ,1～3kb was obtained with cDNA inserted. Conclusions The construction of renal cancer pPC86 yeast two hybrid library would be helpful to renal carcinoma diagnosis and treatment,especially the renal signal treatment.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
2000年第6期325-327,共3页
Chinese Journal of Urology
基金
国家自然科学基金!资助 (39870 841 )
