作　　者： ; ; ; ; ; ;

机构地区： 山西医科大学

出　　处： 《中国生物工程杂志》 2004年第5期73-77,共5页

摘　　要： 为了提高人血小板因子 4(humanplateletfactor 4,hPF4)的表达 ,在PT7 7 hPF4表达质粒的基础上 ,采用PCR定位突变技术 ,改造人血小板因子 4(hPF4)cDNA基因片段 ,去除cDNA 3′端非翻译区AT富含序列 ,改用大肠杆菌强串联终止密码子TAATAA ,成功构建了高效表达质粒pBV2 2 0 hPF4。摇瓶发酵重组人血小板因子 4的产量达 1 60mg L较原表达质粒PT7 7 hPF4表达量提高了近 80倍。经包涵体的洗涤、变性、复性后 ,采用鸡胚绒毛尿囊膜血管生成抑制实验测定复性后rhPF4的生物学活性 ,结果显示 :rhPF4具有抑制血管生成活性。 In order to improve the expression of human Platelet Factor 4 (hPF4) in Escherichia coli , a prokaryotic expression vector pBV220 hPF4 was constructed by DNA polymerase chain reaction(PCR) and DNA recombinant technology,3′ UTR of hPF4 cDNA was deleted and TAG was mutated to TAATAA.The yield of recombinant hPF4 is 160mg /L in shaking flask culture.The expression level has been improved by 80 fold compared with that of PT7 7 rhPF4 expression system.After the inclusion bodies were washed,dissolved and renatured ,inhibition experiment of blood vessel proliferation in chicken chorioallantoic membrane was carried out to determine the bioactivities of rhPF4.The experimental result demonstrated that rhPF4 prepared by this methods had the inhibitory activity against angiogenesis.;

关 键 词： 人血小板因子 大肠杆菌 表达 活性 质粒 血管生成抑制

领　　域： [生物学]