家蚕马氏管免疫相关蛋白基因BmNOS的研究  

Innate immune related gene BmNOS in Malpighian tubule from the silkworm,Bombyx mori

作　　者：唐顺明[1,2] 丁丽平[1] 吴俊[1] 王娜[1] 裘智勇[1,2] 沈兴家[1,2] 

机构地区：[1]江苏科技大学蚕业研究所,江苏镇江212018 [2]中国农业科学院蚕业研究所农业部蚕桑遗传改良重点开放实验室,江苏镇江212018

出　　处：《江苏科技大学学报（自然科学版）》2011年第6期610-615,共6页Journal of Jiangsu University of Science and Technology:Natural Science Edition

基　　金：国家重点基础研究发展计划"973"资助项目(2005CB121000);江苏省高校自然科学基金资助项目(08KJB230001)

摘　　要：借助蛋白双向电泳(2-D)技术对感染家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus,BmNPV)不同时相的家蚕幼虫马氏管蛋白进行分离,并对其中14个差异蛋白点进行质谱分析.结果表明:10个蛋白点得到成功鉴定,其中的一个可信结果为家蚕一氧化氮合酶(Bombyx mori Nitric oxide synthase,BmNOS).由于诱导型BmNOS是合成巨噬细胞免疫效应因子NO的一种关键特异性蛋白酶,因此通过体内注射BmNPV和脂多糖(lipopolysaccharide,LPS)两种诱导剂,采用半定量和实时荧光定量PCR的方法,对感染组和对照组马氏管中BmNOS不同感染时期的表达进行了定量分析.结果表明:经两种诱导剂处理3 h后,BmNOS表达量均迅速递增,BmNPV侵染组上调约7倍;LPS处理组表达量上调25倍,且该趋势持续至6 h,随后逐渐降低并趋于平稳,这表明BmNOS在家蚕马氏管的免疫中具有重要的功能.In order to investigate immune function of Malpighian tubule （MT） in silkworm Bombyx mori, the proteins of MT from larvae infected with Bombyx mori nucleopolyhedrovirus （BmNPV） are separated by using two-dimensional polyacrylamide gel electrophoresis （2-D）. Fourteen protein spots that increase dramatically in the infected ones are excised and identified by means of mass spectrometry. The result shows that 10 spots are i- dentified successfully, and one protein spot has a good match with Bombyx mori nitric oxide synthase （BmNOS）. Inducible nitric oxide synthase is the key one for NO, which is an important effector molecule of maerophages. Moreover, the expression level of BmNOS gene in MT is investigated by quantitative real-time PCR method at dif- ferent development stages of silkworm treated with BmNPV or lipopolysaccharide （LPS）. The result shows that expression level of BmNOS gene is dramatically increased by 7 times at 3 hr post infection of BmNPV, and by 25 times from 3 hr to 6 hr post treatment of LPS, compared with that of the respective control. Hence the expression levels of the control and treatment are similar. The above results suggest that BmNOS may play an important role in innate immune response of silkworm MT.

关 键 词：家蚕 内生免疫 双向电泳 马氏管 家蚕一氧化氮合酶基因 

分 类 号：S881.24[农业科学—特种经济动物饲养]