山西农业大学学报（自然科学版）2019，Vol.39Issue(5)：74-81,8.DOI:10.13842／j.cnki.issn1671-8151.201901030
熊蜂抗冻蛋白基因序列分析及表达特性研究
Sequencing analysis and expression characteristcs of AFP gene in Bombus terriestris
摘要
Abstract
[Objective]T he objective of this study was to analyze the sequence properties of A FP gene from Bombus ter-riestris ,and to investigate its expression patterns at different developmental stages ,tissues and cold stresses .[Meth-ods]Physiochemical properties and structure characteristics of BtAFP were predicted using multiple bioinformatics ap-proaches .T he expression levels of BtA FP mRNA at different developmental stages ,tissues and under different cold stresses of B .terriestris were performed by qRT-PCR .[Results]T he open reading frame (ORF) of BtA FP is 1 143 bp ,encoding a protein of 380 amino acids with an estimated molecular mass of 38 kDa .BtAFP possesses a signal pep-tide and no transmembrane domains .α-helix was an important part of the secondary and tertiary structures of BtAFP , w hich was similar to the structure of fish typeI AFPs .A promoter sequence and several transcription factors associated with environmental changes were identified in 5’U T R region of BtA FP .Phylogenetic tree analysis showed that BtAFP has 74% amino acid sequence identify and the closest evolutionary relationship with the antifreeze proteins of Apidae . T he result of qRT-PCR revealed that the BtA FP transcripts were differentially expressed in different tissues and devel-opmental stages of B .terriestris (P＜0.05).Pupal with brow n eyes had the highest expression level of BtA FP .T he expression profiles of BtA FP in antenna ,wings and legs of adult workers were higher than that in other tissues .Under cold stresses ,the mRNA expression of BtA FP was changed by the increase of process time .T he expression level of BtA FP at －20℃for 20 min and 0 ℃ for 16 h were significantly higher than other given time ,respectively .[Conclu- sion]T hese result suggested that BtA FP plays an essential role in grow th and response to cold stresses of Bombus ter-riestris ,w hich might be used as a candidate gene for further studies on cold resistance of bumble bees .关键词
熊蜂/抗冻蛋白/发育表达/低温胁迫Key words
Bombus terriestris/Antifreeze protein/Developmental expression/Low temperature stresses分类
农业科技引用本文复制引用
Xu Kai,Chen Donghai,Wang Zhi,Liu Yuling,Wang Zhenwei,Niu Ben,Cai Xiangqiang,Niu Qingsheng..熊蜂抗冻蛋白基因序列分析及表达特性研究[J].山西农业大学学报（自然科学版）,2019,39(5):74-81,8.基金项目
国家蜂产业技术体系专项(CARS-44-SYZ 4) （CARS-44-SYZ 4）
吉林省中青年科技创新领军人才及团队项目(20180519009JH ) （20180519009JH ）
吉林省科技创新中心 －基地与科技服务平台建设(20180623003T C) （20180623003T C）
吉林省重点科技攻关项目(20160204014NY ) （20160204014NY ）
