安徽农业科学2018，Vol.46Issue(12)：112-117,6.
米虾鞣化激素的表达特征分析及体外合成
Expresstion Characteristics Analysis and in vitro Synthesis of Bursicon from Caridina
摘要
Abstract
[Objective]To observe the process of embryo development in Caridina and express the proteins of bursicon-αand bursicon-β.Be-sides,to explore the expresstion characteristics of bursicon in different tissues from Caridina.[Method]The light microscopy was used to observe the embryo development.According to the transcriptome hints,the coding regions of bursicon-αand bursicon-βwere amplified with PCR.The PCR products were inserted into the pET-28a-c(+)vector after double digestion to construct expresstion vectors,pET-28a-αand pET-28a-β.The re-combinant plasmids were transformed into Rosetta and expressed by induction with IPTG.The expression differences of bursicon-αand bursicon-β in four tissues of eyestalk,heart,thoracic ganglia and abdominal ganglia from Caridina were detected by the real-time quantitative PCR.[Result] The process of embryo development in Caridina included seven stages:cleavagestage,blastula,gastrula,egg-nauplius stage,egg-metanauplius stage,embryo with eye pigments formed stage and prehatching stage.Bursαand Bursβwere obtained successfully by prokaryotic expression ana-lyzed by SDS-PAGE.The relative molecular mass of both bursicon subunits were 15 kD.In addition,the expresstion characteristics analysis showed that the relative expression levels of bursicon-αand bursicon-βwere reached to the highest level in abdominal ganglia and declined in thoracic ganglia.Nevertheless,low level relative expressions were clearly detectable in the eyestalk and heart.[Conclution]These findings indicated that bursicon-αand bursicon-βexpressed mainly in abdominal ganglia.Bursicon is a kind of neuropeptide hormone.It plays a regulatory role by relea-sing into hemolymph.The proteins of Bursαand Bursβobtained by prokaryotic expression laid a foundation of further study on the function.关键词
米虾/鞣化激素/原核表达/组织表达/实时荧光定量PCRKey words
Caridina/Bursicon/Prokaryotic expression/Tissue expression/Real-time quantitative PCR分类
农业科技引用本文复制引用
任丽琦,翁洁羊,王鑫,孙金生,李冉..米虾鞣化激素的表达特征分析及体外合成[J].安徽农业科学,2018,46(12):112-117,6.基金项目
国家自然科学基金青年基金项目(31302168) （31302168）
天津市应用基础与前沿技术研究计划(14JCYBJC30700) （14JCYBJC30700）
天津师范大学博士基金项目(52XB1303). （52XB1303）
