家蚕胰脂肪酶相关蛋白1基因(BmPLRP1)的克隆及表达分析  

Cloning and expression of BmPLRP1 gene in pancreatic lipase-related protein 1 of silkworm,Bombyx mori

作　　者：李玉霞 李刚[1,2] 钱荷英 郝长富[1] 刘明珠 赵国栋 徐安英[1,2] LI Yuxia;LI Gang;QIAN Heying;HAO Changfu;LIU Mingzhu;ZHANG Guodong;XU Anying(Jiangsu University of Science and Technology,Zhenjiang 212018,China;The Sericultural Research Institute,Chinese Academy of Agricultural Sciences,Zhenjiang 212018,China)

机构地区：[1]江苏科技大学生物技术学院,江苏镇江212018 [2]中国农业科学院蚕业研究所,江苏镇江212018

出　　处：《河北农业大学学报》2020年第1期82-88,共7页Journal of Hebei Agricultural University

基　　金：江苏省自然科学基金面上项目(BK20181228);现代农业产业技术体系建设专项(CARS-18);镇江市重点研发计划(现代农业)项目(NY2018021,NY2017017).

摘　　要：胰脂肪酶相关蛋白1(Pancreatic lipase-related protein 1,PLRP1)是由PLRP1基因编码的一种脂肪酶,在通常状态下无活性。PLRP1的分泌与生物的生长发育有着密切的联系,但该基因在家蚕中的功能尚未清楚。本研究克隆获得家蚕PLRP1基因(BmPLRP1)的核苷酸序列,并对其序列特征进行生物信息学分析,进一步采用半定量RT-PCR和荧光定量PCR检测BmPLRP1基因的表达水平。结果表明,该基因位于家蚕第12号染色体的nscaf2993位点,编码331个氨基酸,等电点为8.72,预测的BmPLRP1蛋白无信号肽、分子质量为37.1 kD,没有跨膜结构域,具有一个Pancreatlipaselike结构域,存在活性位点、磷酸化位点、O-糖基化位点,不存在N-糖基化位点。系统进化分析表明BmPLRP1与野桑蚕的PLRP1的进化关系较近;RT-PCR检测表明,BmPLRP1基因在家蚕5龄3天幼虫的头部、表皮、丝腺、生殖器、气管丛、中肠中有明显表达,而在血液和脂肪体中的表达量相对较低;对家蚕品种‘733xin’喂食经100 mg/kg NaF溶液浸泡过的桑叶后,实时定量PCR检测BmPLRP1基因在中肠、血液、丝腺、脂肪体中的相对表达量,结果表明均有所上调,推测该基因可能参与了氟化物侵染家蚕后的应答反应过程。Pancreatic lipase-related protein 1(PLRP1)is a lipase encoded by the PLRP1 gene and is inactive under normal conditions.The secretion of PLRP1 is closely related to the growth and development of organisms;however,the function of PLRP1 in silkworm is still unclear.In this study,we acquired the nucleotide sequence of the BmPLRP1 gene.The expression profile of BmPLRP1 was further detected by semi-quantitative RT-PCR and real-time PCR.The results showed that BmPLRP1 is located on nscaf2993 of silkworm chromosome 12,encoding 331 amino acids with an isoelectric point of 8.72.The predicted BmPLRP1 protein has no signal peptide,and the molecular mass is 37.1kD,having no transmembrane domain,but a Pancreat_lipase-like domain.The domain has an active site,a phosphorylation site,an O-glycosylation site,and no N-glycosylation site.Phylogenetic analysis indicated that BmPLRP1 has a close relationship with PLRP1 of Bombyx mandarina;RT-PCR analysis showed that the BmPLRP1 gene was expressed in the head,epidermis,silk gland,genital,tracheal plexus and midgut of 5th instar larvae of silkworm,but the expression level in blood and fat body was relatively lower.After induction of silkworm cultivars 733xin by mulberry leaves soaked by 100 mg/kg NaF,the relative expression of BmPLRP1 gene in the midgut,blood,silk gland and fat body was up-regulated.This gene may be involved in the response process after fluoride infects the silkworm.

关 键 词：家蚕 胰脂肪酶相关蛋白1(PLRP1) 克隆 序列特征 表达谱 

分 类 号：S881.26[农业科学—特种经济动物饲养]