葡萄胎病理相关新基因F10重组蛋白的表达及鉴定  

Expression and identification of recombinant protein of the new gene F10 which is associated with the pathogenesis of hydatidiform mole

作　　者：庞战军[1] 周君桂[2] 

机构地区：[1]南方医科大学南方医院妇产科,广东医学博士510515 [2]南方医科大学南方医院康复理疗科,广东510515

出　　处：《医学研究生学报》2014年第6期577-581,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81170595)

摘　　要：目的 F10是从葡萄胎组织中克隆的新基因,前期研究显示F10基因可能与滋养细胞侵袭性有关,为进一步研究F10基因功能,拟构建原核质粒表达葡萄胎相关新基因F10重组蛋白,分离纯化并鉴定。方法用逆转录聚合酶链反应法从人组织中扩增F10基因编码序列,将其克隆到pET28a原核表达载体,在大肠杆菌BL21(DE3)内进行诱导表达F10基因重组蛋白,用层析柱予以纯化并采用SDS-PAGE电泳鉴定。结果成功构建了pET28a/F10原核表达质粒,并获得高纯度的重组F10蛋白。结论表达及纯化的重组F10蛋白纯度高,适合作为制备单克隆抗体的抗原,可用于F10功能的进一步研究。Objective F10 is a new gene cloned from hydatidiform moles.Our previous study showed that the F10 gene might be associated with the invasion of trophoblast cells.This study was to construct a prokaryotic plasmid expressing the recombinant protein of F10 and isolate,purify,and identify it in order to further investigate the function of the F10 gene.Methods The coding sequence of the F10 gene was amplified from the human tissue by RT-PCR and cloned into the vector pET28a.The plasmid was expressed in the E.coli BL21（ DE3） to obtain the F10 recombinant protein.The expressed F10 recombinant protein was purified with the chromatographic column and verified by sodium dodecyl sulfate polyacrylamide gel electropheresis（ SDS-PAGE）.Results The prokaryotic plasmid expressing the F10 recombinant protein was successfully constructed and highly purified F10 recombinant protein was obtained.Conclusion The F10 recombinant protein was expressed and highly purified,which could be used as an antigen for the preparation of the F10 monoclonal antibody.

关 键 词：F10基因 F10重组蛋白 葡萄胎 

分 类 号：R737.33[医药卫生—肿瘤]