利用在大肠杆菌体内表达的重组小麦组蛋白H4与质粒DNA形成复合体后,通过琼脂糖凝胶电泳迁移试验(EMSA)验证重组蛋白与DNA的结合情况;MTT法检测蛋白对细胞的毒性作用;转染细胞后,激光共聚焦显微镜检测荧光蛋白的表达。此重组蛋白能很好地结合质粒DNA,且能有效地保护DNA分子不受核酸酶降解,并能高效地携带绿色荧光蛋白基因pEGFP/C1转染进入卵巢癌细胞株H08910。
Recombinant wheat hisone H4 was expressed in E.Coli. Purified protein binded plasmid DNA using gel electrophoresis mobility shift assay (EMSA).Cell proliferation was measured with MTT assay.Green fluorescent protein was measured using a confocal microscope.This recombinant wheat histone H4 could bind plasmid DNA efficiently and protect the DNA from DNase I.Plasmid pEGFP/C1 was transfected into ovarian cancer cells HO8910 and obtained expression with high efficacy.