摘要
目的在大肠埃希菌中表达磷脂氢谷胱甘肽过氧化物酶(phospholipidhydroperoxideglutathioneperoxidase,PHGPx),以期用于抗PHGPx抗体的制备。方法用特异引物从基因文库中扩增出PHGPxcDNA,与表达载体重组,转化大肠埃希菌。重组体中的PHGPx经点突变作用,编码硒半胱氨酸的密码子UGA变为编码半胱氨酸的UGU。结果序列分析表明,克隆载体中包含PHGPx的完整编码序列及3'未翻译区序列;经SDS鄄PAGE及WesternBlot证实PHGPx在大肠埃希菌中获得了成功表达。结论突变的PGPGx可以在大肠埃希菌中高效表达并可用于制备抗PHGPx抗体。
Objective To express human phospholipid hydroperoxide glutathione peroxidase(PHGPx) in Escherichia coli ( E.coli ). Methods The cDNA of PHGPx was amplified from a human testis library using specific primers and cloned into the expression vector pRSET. The selenocysteine codon UGA in PHGPx was mutated into cysteine codon UGU. E.coli was transformed by the recombinant structure. Results The entire structure gene and the 3' UTR (untranslated region ) of PHGPx were cloned into the cloning vector and verified by sequencing. Conclusion The mutated human PHGPx is expressed in E.coli and it could be used as antigen in the preparation of anti-PHGPx antibody.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
2004年第2期119-120,共2页
Chinese Jouranl of Endemiology
基金
国家自然科学基金资助项目(30170803)
黑龙江省科技攻关资助项目(GB01C10601)
