摘要
将美洲拟鲽抗冻肽基因克隆于原核高效表达载体ｐＫＫ２２３－３中，经酶切分析、ＰＣＲ检测筛选出重组克隆ｐＭＫ１１，转化大肠杆菌ＪＭ１０９和ＤＨ５α，ＩＰＴＧ诱导２～３ｈ后，超声波裂解细菌产物，在ＳＤＳ－ＰＡＧＥ电泳图谱分子量９ｋＤａ处显示出一条明显的蛋白带，与美洲拟鲽抗冻肽大小一致，表明美洲拟鲽抗冻肽基因在大肠杆菌中表达，为进一步研究抗冻肽基因在大肠杆菌中的表达水平及条件打下基础．
In order to immunological assay for antifreeze peptide (AFP) from transgenic tobacco plants,AFP should be purified as an antigen,and AFP antibody should be prepared.An EcoRI/HindⅢ fragment containing AFP coding gene is subcloned into the bacterial expression vector pKK223-3.The protein pattern on SDS-PAGE indicated that 9 kDa protein band as the same as flounder AFP preproprotein is induced by the addition of IPTG to the exponential phase culture of E.coli harboring AFP expression recombinant plasmid pMK11.
出处
《内蒙古大学学报（自然科学版）》
CAS
CSCD
1999年第2期236-238,共3页
Journal of Inner Mongolia University：Natural Science Edition
基金
国家自然科学基金
