CAJ | 학술논문

目的通过昆虫-杆状病毒表达系统表达获得Legumain蛋白,为进一步研究其在葡萄膜黑色素瘤中的作用建立基础.方法实验研究.对2011年4月至2012年1月在陕西省眼科中心应用克隆Legumain蛋白基因,并将其基因片段插入pFastBac载体获得重组载体,转化大肠杆菌DH10Bac感受态细胞后得到重组Bacmid,转染昆虫Sf9细胞,Western Blotting印迹和SDS-PAGE检测重组蛋白的表达.结果获得了Legumain蛋白的重组杆状病毒;Western Blotting印迹和SDS-PAGE分析表明,该重组杆状病毒感染昆虫Sf9细胞后表达Legumain蛋白,与预期结果一致.结论通过昆虫-杆状病毒表达系统可成功获得Legumain蛋白,从而为进一步研究其在葡萄膜黑色素瘤侵袭与转移机制的作用奠定了工作基础.
목적 통과곤충-간상병독표체계통표체획득Legumain단백,위진일보연구기재포도막흑색소류중적작용건립기출.방법 실험연구.대2011년4월지2012년1월재합서성안과중심응용극륭Legumain단백기인,병장기기인편단삽입pFastBac재체획득중조재체,전화대장간균DH10Bac감수태세포후득도중조Bacmid,전염곤충Sf9세포,Western Blotting인적화SDS-PAGE검측중조단백적표체.결과 획득료Legumain단백적중조간상병독;Western Blotting인적화SDS-PAGE분석표명,해중조간상병독감염곤충Sf9세포후표체Legumain단백,여예기결과일치.결론 통과곤충-간상병독표체계통가성공획득Legumain단백,종이위진일보연구기재포도막흑색소류침습여전이궤제적작용전정료공작기출.
Objective To obtain human Legumain protein with insect-baculovirus expression system,make foundation for the uveal melanoma study.Methods The gene encoding Legumain protein was cloned and inserted into the pFastBac vector,and the recombinant bacmid was generated after the E.coli DH10Bac competent cell was transformed by the recombinant pFastBac vector.The recombinant baculovirns was transfected into insect cell Sf9.Western Blotting and SDS-PAGE analysis of recombinant protein expression was performed.Results Recombinant baculovirns expressing Legumain protein was obtained.Western Blotting and SDS-PAGE analysis showed that Legumain protein was expressed when the Sf9 cells were infected by the recombinant baculovirus,which consistent with the expected results.Conclusions Legumain protein was obtained through insect-baculovirus expression system.Cloning and expression of Legumain protein provide the foundation for the mechanism of Legumain in the uveal melanoma invasion and metastasis.