摘要
目的 建立酵母双杂交系统 ,筛选与FasL相互作用的蛋白 ,探讨FasL与大肠癌肝转移的关系。方法 以FasL基因构建诱饵蛋白质粒 ,筛选人胎肝cDNA文库 ,鉴定与FasL相互作用的蛋白 ,通过生物信息学分析FasL及其相互作用蛋白在大肠癌肝转移中的作用。结果 筛选出 10个与FasL特异性相互作用的蛋白 ,包括金属硫蛋白 1K、1G、2A ,组织蛋白酶B ,脂肪酸合成酶 ,干扰素α诱导蛋白 2 7,磷脂清除酶 ,丝氨酸 /苏氨酸样激酶 ,锚着黏附蛋白以及纤维微丝蛋白 5等。结论 成功建立了筛选FasL相互作用蛋白的酵母双杂交系统 ,并初步证明FasL与组织蛋白酶、金属硫蛋白。
Objective To construct the yeast two hybrid system, and screen the proteins which interact with FasL, and investigate the relationship of FasL and hepatic metastasis of colorectal carcinoma. Methods We have cloned the FasL gene into the pGBKT7 vector as the bait, then screened the fetal liver cDNA library, and have got a series of specific proteins that interact with FasL protein. Using the bioinformatics, we analyzed the interacting proteins in the mechanism of hepatic metastasis of colorectal carcinoma. Results We have screened several proteins that interaction with FasL protein, including metallothionein 1K、1G、2A, cathepsin B, fatty acid synthase, interferon alpha inducible protein 27, phospholipid scramblase, Ser/ Thr like kinase, anchor attachment protein,fibulin 5.Conclusions We have successfully constructed the yeast two hybrid system, and preliminary identified that the interaction between FasL, metallothionein, cathepsin and anchor attachment protein is radically related to the hepatic metastasis of colorectal carcinoma.
出处
《中华外科杂志》
CAS
CSCD
北大核心
2004年第11期672-674,共3页
Chinese Journal of Surgery
基金
国家自然科学基金资助项目 (3 0 0 70 747)
关键词
蛋白质
酵母双杂交系统
大肠癌
肝转移
Yeast
Two-hybrid system techniques
Colorectal neoplasms
Neoplasm metastasis
