家蚕脂肪酶1(Bmlipase-1)的原核表达和抗体制备及在转基因增量表达系统中的检测  

Prokaryotic Expression and Antibody Preparation of Bmlipase-1 and Determination of Overexpression in Transgenic Silkworm

作　　者：金盛凯[1] 蒋亮[1] 林平[1] 孙薇[1] 程廷才[1] 夏庆友[1] 

机构地区：[1]西南大学家蚕基因组生物学国家重点实验室,重庆400715

出　　处：《蚕业科学》2012年第6期1000-1004,共5页ACTA SERICOLOGICA SINICA

基　　金：国家重点基础研究发展计划"973"项目(No.2012CB1146-00);国家高技术研究发展计划"863"项目(No.2011AA-100306)

摘　　要：家蚕肠液中的脂肪酶1(Bmlipase-1)对家蚕核型多角体病毒(BmNPV)具有明显抑制作用,建立的增量表达Bmlipase-1的转基因家蚕品系(LI-A)对BmNPV的抵抗力得到显著增强。为了分析LI-A品系幼虫肠液中Bmlipase-1的含量变化,通过原核表达、纯化Bmlipase-1蛋白,并免疫家兔制备Bmlipase-1的多克隆抗体,对LI-A品系和正常家蚕品种大造4龄、5龄起蚕肠液中的Bmlipase-1含量进行Western blotting检测。结果显示Bmlipase-1在LI-A品系幼虫肠液中的含量明显高于正常家蚕品种大造,证明LI-A品系对BmNPV的抵抗力提高是因为肠液中Bmlipase-1的含量增加。Bombyx mori lipase 1（Bmlipase-1） existing in larval intestinal juice has a obvious antiviral activity against Bombyx mori nucleopolyhedrovirus（BmNPV）.We previously demonstrated that the resistance to BmNPV of transgenic silkworm lineage LI-A overexpressing Bmlipase-1 was significantly enhanced.In an attempt to analyze the content variation of Bmlipase-1 in digestive juice of LI-A,we obtained Bmlipase-1 protein via prokaryotic expression.After purification,Bmlipase-1 was used to immunize rabbit for preparing polyclonal antibody.Furthermore,we determined Bmlipase-1 content in intestinal juice of the 4th and 5th instar newly exuviated larvae of LI-A and normal silkworm variety Dazao by Western blotting.The results revealed that the content of Bmlipase-1 in the digestive juice of LI-A was significantly higher than that of Dazao,demonstrating that the enhanced anti-BmNPV activity of LI-A was due to the increase of Bmlipase-1 content in the intestinal juice.

关 键 词：家蚕核型多角体病毒 脂肪酶1 原核表达 多克隆抗体 转基因 抵抗力 家蚕 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]