九香虫脂肪酸合酶基因克隆、生物信息学分析及时序表达谱研究  

Cloning,Bioinformatics Analysis and Temporal Expression of the Fatty Acid Synthase Gene in Aspongopus chinensis Dallas

作　　者：冯芷汀 尹志勇 郭建军[1] Feng Zhiting;Yin Zhiyong;Guo Jianjun(Institute of Entomology,Guizhou University/Guizhou Provincial Key Laboratory for Agricultural Pest Management of the Mountainous Region,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州大学昆虫研究所/贵州省山地农业病虫害重点实验室,贵州贵阳550025

出　　处：《山地农业生物学报》2024年第3期15-20,共6页Journal of Mountain Agriculture and Biology

基　　金：国家自然科学基金(82160743)。

摘　　要：九香虫Aspongopus chinensis Dallas,1851是中国传统药食两用资源昆虫,其需求量日增。然其存在严格的生殖滞育现象,脂肪酸合酶(Fatty Acid Synthase,FASN)与昆虫滞育关系密切。本文根据九香虫转录组数据设计了FASN特异性引物,提取九香虫总RNA,利用PCR技术克隆出九香虫脂肪酸合酶基因(AcFASN),利用生物信息学软件对其进行了生物信息学和同源进化树分析,并通过实时荧光定量试验获取了AcFASN在九香虫中的时序表达图谱。结果显示:九香虫AcFASN基因全长7092 bp,编码2363个氨基酸;九香虫AcFASN是亲水性蛋白质,亚细胞主要定位于细胞质,是一种无信号肽、非跨膜、非分泌型蛋白。同源进化树分析表明,AcFASN氨基酸序列与茶翅蝽Halyomorpha halys的FASN氨基酸序列进化关系较近。时序表达谱显示AcFASN基因在滞育期表达量较高,预示着AcFASN基因极可能通过改变九香虫体内AcFASN的表达量来影响九香虫滞育等生命活动。Aspongopus chinensis Dallas,1851,is a traditional medicinal and edible insect in China,with an increasing demand.Nevertheless,it shows strict reproductive diapause,which is influenced by Fatty Acid Synthase(FASN).In this study,specific primers were designed for FASN based on the transcriptome data of A.chinensis,and total RNA was extracted and cloned using PCR.The FASN gene,known as AcFASN,was successfully cloned through the process of RNA extraction,reverse transcription,PCR,and ligation transformation.Bioinformatics and homologous evolutionary tree analysis were conducted using the gene sequence of FASN,while the temporal expression graphs of AcFASN were generated through real-time fluorescence quantitative PCR.The purpose of these analyses was to determine if AcFASN plays a role in diapause regulation.The results revealed that the AcFASN gene spans a total length of 7092 bp and codes for 2363 amino acids.Bioinformatics analysis showed that AcFASN was a hydrophilic protein that was non-secreted and was found in the cytoplasm without a transmembrane region or signal peptide.Phylogenetic tree analysis revealed a strong evolutionary connection between the FASN amino acid sequences of A.chinensis and Halyomorpha halys.Temporal expression analysis showed that the AcFASN expression level was elevated during diapause,suggesting that AcFASN may play a role in regulating the growth and development of A.chinensis by modulating its expression in the body.

关 键 词：脂肪酸合酶 九香虫 基因克隆 时序表达谱 

分 类 号：S899.9[农业科学—特种经济动物饲养]