琥珀蚕胆绿素结合蛋白基因AaBP-1的克隆及表达分析  

Cloning and Expression Analysis of Biliverdin-binding Protein Gene (AaBP-1) from Antheraea assama

作　　者：李琼艳[1] 陈安利[1] 钟健[1] 刘增虎[1] 廖鹏飞[1] 杨伟克[1] 董占鹏[1] Li Qiongyan;Chen Anli;Zhong Jian;Liu Zenghu;Liao Pengfei;Yang Weike;Dong Zhanpeng(Institute of Sericulture and Apiculture,Yunnan Academy of Agricultural Sciences,Mengzi Yunnan 661101,China)

机构地区：[1]云南省农业科学院蚕桑蜜蜂研究所

出　　处：《蚕业科学》2019年第3期353-360,共8页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目（No.31360586）;云南省应用基础研究项目（No.2017FB069）

摘　　要：鳞翅目昆虫的胆绿素结合蛋白属脂质运载蛋白家族,通过与胆色素(bilins)结合在蓝绿色素形成过程中发挥重要作用。从琥珀蚕(Antheraea assama)丝腺中克隆了胆绿素结合蛋白基因1(AaBP-1),该基因cDNA全长923 bp,开放阅读框609bp,编码202个氨基酸残基,预测蛋白质分子质量为22.1 kD,理论等电点为5.19,蛋白序列含有1个信号肽和1个Lipocalin超家族结构域。序列比对分析结果表明,AaBP-1的氨基酸序列与柞蚕胆绿素结合蛋白BP-1一致性较高,达到81.7%。荧光定量PCR检测结果显示AaBP-1基因mRNA在琥珀蚕5龄第4天幼虫的表皮和丝腺中高量表达,在头和中肠中少量表达,其他组织中都几乎不表达。在丝腺中,AaBP-1主要在前部丝腺和中部丝腺表达,尤其在中部丝腺的前部大量表达,且琥珀蚕丝腺的颜色也呈现出中部丝腺比后部丝腺深的现象。推测AaBP-1可能参与琥珀蚕表皮、丝腺及茧中色素的形成。研究结果将为进一步研究琥珀蚕色素形成机制提供参考。As a member of lipocalin protein family,biliverdin-binding proteins of lepidopteran insects play an important role in the formation of blue-green pigment through binding bilins. In this study,a 923 bp full length cDNA of biliverdinbinding protein gene(AaBP-1)was cloned from the silk gland of Antheraea assama. The open reading frame was 609 bp,encoding 202 amino acid residues. The predicted molecular weight of AaBP-1 protein was 22 1 kD and the theoretical isoelectric point was 5 19. The protein contained a signal peptide and a Lipocalin superfamily domain. Sequence alignment analysis showed that the amino acid sequence of AaBP-1 shared 81 7% similarity to biliverdin-binding protein BP-1 of Antheraea pernyi. qRT-PCR analysis indicated that the expression level of AaBP-1 mRNA was high in epidermis and silk gland of Antheraea assama larvae of 4th day 5th instar,low in head and midgut,almost none in other tissues.Furthermore,AaBP-1 was mainly expressed in anterior and middle silk gland,especially in anterior part of middle silk gland. The color of the middle silk gland was deeper than that of the posterior silk gland. It was speculated that AaBP-1 might be involved in pigment formation in epidermis,silk gland and cocoon of Antheraea assama. The results provide a reference for further study on mechanism of pigment formation in Antheraea assama.

关 键 词：琥珀蚕 胆绿素结合蛋白 基因克隆 序列分析 表达谱 

分 类 号：S885.9[农业科学—特种经济动物饲养] Q786[农业科学—畜牧兽医]