CAJ | 학술논문

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。
목적： F10시종포도태조직중극륭적신기인，전기연구현시F10기인가능여자양세포침습성유관，위진일보연구F10기인공능，의구건원핵질립표체포도태상관신기인 F10중조단백，분리순화병감정。방법용역전록취합매련반응법종인조직중확증F10기인편마서렬，장기극륭도pET28a원핵표체재체，재대장간균BL21（DE3）내진행유도표체F10기인중조단백，용층석주여이순화병채용SDS-PAGE전영감정。결과성공구건료pET28a/F10원핵표체질립，병획득고순도적중조F10단백。결론표체급순화적중조F10단백순도고，괄합작위제비단극륭항체적항원，가용우F10공능적진일보연구。
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 pET 28a.The plasmid was ex-pressed 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 pro-karyotic plasmid expressing the F 10 recombinant protein was successfully constructed and highly purified F 10 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 .