人α、β珠蛋白基因以串联形式在大肠杆菌中的表达  

Serial Expression of Human α,β globin Genes in Escherichia coli

作　　者：张浩[1] 王全立 冯起 杜芝燕 李晓霞[3] 毛秉智[1] 

机构地区：[1]北京放射医学研究所,北京100850 [2]北京输血医学研究所 [3]解放军总医院临床检验科

出　　处：《中国输血杂志》2001年第2期65-67,共3页Chinese Journal of Blood Transfusion

摘　　要：目的　为开展以基因重组血红蛋白为基础的血液代用品研究 ,尝试将人α、β珠蛋白基因以串联形式在大肠杆菌中进行表达。方法　采用常规的分子生物学技术。结果　用PCR扩增出含血红蛋白α、β珠蛋白的串联基因片段 ,测序表明克隆的α、β基因未发生意外突变。将该串联基因克隆进 pBV2 2 0表达载体 ,经热诱导表达产物达细菌总蛋白的 2 0 %左右。该表达产物以包涵体形式存在 ,并经WesternBlotting证实。Objective To make an attempt to express serialy the human α,β globin in Escherichia Coli in order to carry out the research on blood substitute based on the genetic recombination of hemoglobin.Methods The routine molecular biology technology was used.Results The serial genetic fragment of α,β globins containing the hemoglobin was amplified by PCR.The genetic sequencing showed that there was no unexpected mutation.And then,the serial genes were cloned into the pBV220 expression carrier.Undergoing thermal inducement,the expression product could reach about 20% of total bacterial protein.The expression product assumed the form of inclusion body,and it was vertified by Western Blotting.Conclusion This much can lay the solid foundations for further launching the research on expression and pruification of recombinant human hemoglobin genes.

关 键 词：血红蛋白 大肠杆菌 基因表达 α β珠蛋白 

分 类 号：R378.21[医药卫生—病原生物学]