摘要
目的 :探索重组人血红蛋白的纯化方法。方法 :将α、β珠蛋白串联基因克隆进 pBV2 2 0表达载体 ,获得了高效表达 ,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 ,包涵体经洗涤后 ,用 8mol/L尿素溶解 ,先用Q SepharoseFastFlow阴离子交换纯化后 ,又经Sephacryl 10 0凝胶过滤纯化。 结果 :经两步纯化后重组人血红蛋白的纯度达 90 %左右。纯化产物经复性 ,具有与氧结合的能力。结论
Purpose:The aim is to study the purification methods of recombinant human hemoglobyin.Methods:Conjugated α and β globin chain genes were introduced into expression vector pBV220. The expression product reached about 20% of total bacterial protein. They were expressed as inclusion bodies.After washing with 4 mol/L urea, inclusion bodies were dissolved in 8 mol/L urea.Purification was first performed on a column of Q sepharose Fast Flow,and further purification was performed on a column of Sephacry 100,which resulted in the elimination of most of the contaminated proteins.Results:The purity of recombinant human hemoglobin is about 90% after two steps of purification. The purification product had biological activity after refolding and renatured reaction.Conclusions:The methods to purify recombinant human hemoglibin were established.
出处
《中国生化药物杂志》
CAS
CSCD
2000年第6期274-277,共4页
Chinese Journal of Biochemical Pharmaceutics
关键词
血红蛋白
大肠杆菌
表达
蛋白质纯化
Hemoglobin
Escherichia coli
Experssion
Protein purification
