遗传 ›› 1993, Vol. 15 ›› Issue (5): 6-10.
王汝刚1;秦士良2 WANG Ru-Gang1;QIN Shi-Liang
摘要: 本文应用近年发展起来的生化技术――蛋白质双向电泳(其第一向为等电聚焦,第二向为SDS凝胶电泳),将小鼠腹水细胞核糖体蛋进行了指纹分离。并利用蛋白质印迹转移(Western blotting),将转移后的硝酸纤维膜与交联了碱性磷酸酶的第二抗体和抗酵母EF-3抗体反应,证实该核糖体蛋白含有EF-3同源片段,进而制备了蛋白质合成无细胞体系。通过测定PolyU指导下3 H-phe掺入活力的免疫失活实验,初步证实此同源片段是小鼠腹水细胞蛋白质合成所必需。
The ribosomal proteins of H22a cell,were separated with the method of two-D gel electrophoresis (the first dimention is isoelectric focus and the second is SDS PAGE).Then the Western blotting was used,the transferred nitrocellulose sheet was treated with antiyeast EF-3 antibody and the second antibody bonded with alklinephosphoesterase.The result shows that the ribosomal proteins have a homologous fragment to yeast EF-3 factor.The cell-free system of protein synthesis was also established.By determing the activity of polyU direeted 3H-phe intervention in immunodeactivitive experiment,it is primaril confirmed that this fragment is the esscntial for the protein biosynthesis in H22a cell.