摘要
抗菌肽是存在于生物细胞中的一类具有强抗菌作用的天然小分子多肽,它们在机体免疫系统中发挥了重要作用,被认为是抗生素的理想替代品。本文通过RT-PCR法从斑点叉尾鮰(Ictalurus punctatus)的肝脏中分离到抗菌肽-2(LEAP-2)基因,该基因编码94个氨基酸残基组成的多肽;氨基端的信号肽由28个残基组成,成熟肽含有41个残基;对于鱼类和哺乳动物显示高度保守的4个半胱氨酸残基位于多肽的羧基端区域。通过对LEAP-2基因添加EcoRI和HindIII酶切位点和对pET32a进行双酶切处理,成功构建了"pET32a-IpL-2"重组表达质粒,并转化工程菌E.coliBL21(DE3)。在工程菌对数生长后期加入诱导剂IPTG、经25℃培养后,可溶性的"trxA-IpL-2"融合蛋白得到高程度的表达。Tricine-SDS-PAGE和Western blot分析表明,获得了高纯度的目的蛋白。
Antimicrobial peptides generally are some small molecule peptides produced by the cells of living organisms,and they play an important role in the host s immune response against microbial invasion.Thus,they are considered to be good substitutes for traditional antibiotics.In the present study,a liver-expressed antimicrobial peptides 2(LEAP-2) gene was amplified from the liver of channel catfish(Ictalurus punctatus) by RT-PCR.This LEAP-2 open reading frame encoded a peptide consisting of 94 amino acid residu...
出处
《上海交通大学学报（农业科学版）》
2010年第1期70-75,96,共7页
Journal of Shanghai Jiaotong University(Agricultural Science)
