胃癌组织中EphB_2受体胞外区结构域的克隆和序列测定

Cloning and sequencing of the ectodomain of EphB_2 receptor in the tissue of gastric carcinoma

目的 对胃癌组织中酪氨酸蛋白激酶 EphB2 受体胞外区结构域进行克隆。方法 从胃癌患者手术切片标本中提取RNA,进行RT PCR,将扩增片段克隆入 pUC19质粒,进行测序及酶切鉴定。结果 克隆片段序列正确。结论 为进行表达、筛选与之相互作用的蛋白,阐明其生理功能打下基础。

Objective To clone the ectodomain of EphB 2 receptor tyrosine kinase in the tissue of gastric carcinoma.Methods Total RNA was extracted from a surgical specimen of human gastric carcinoma, and primers were designed by computer analysis and chemically synthesized. The amplified fragment was obtained by RT-PCR and cloned into pUC 19 vector. The recombinant clones were confirmed by sequencing and restriction endonuclease cleavages. Results The cloned fragment was identical to the design.Conclusion It will offer the basis for further investigation including identify its new ligand, the expression, and biological functions.