棉铃虫中肠氨基氨肽酶N基因克隆及序列分析.pdfVIP

中国农业科学 　2003 ,36 ( 11) :1293 - 1300 Scientia Agricultura Sinica 棉铃虫中肠氨基氨肽酶 N 基因的克隆及序列分析 王桂荣 ,梁革梅 ,吴孔明 ,郭予元 ( 中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室 , 北京 　100094) 摘要 : 通过简并引物 PCR 结合 RACE 技术 ,从棉铃虫中肠中克隆了 1 个氨基氨肽酶 N 基因 ,该基因全长 3 043 bp ,阅读框包括 2 856 bp ,编码 951 个氨基酸 。预测蛋白分子量为 108. 3 kD ,等电点为 5. 29 。序列中具有谷氨酸锌化 氨肽酶的保守结构 GAMEN 以及锌结合位点 HEXXHX18 E 。N末端 20 个氨基酸具有信号肽的特征 ,C末端 25 个氨基 酸具有明显的亲脂性 ,并且具有跨膜特征 。该基因已经在 GenBank 中登记 ,序列号为 :AY181026 。 关键词 : 棉铃虫 ; 氨基氨肽酶 N ; 基因克隆 Cloning and Se quencing of a Gene Enco ding Aminop eptida se N in the Midgut of Helicoverp a armigera ( Hübner) WAN G Guirong , L IAN G Gemei , WU Kongming , GUO Yuyuan ( State Key L aboratory of Plant Disease and Insect Pests , Institute of Plant Protection , Chinese Academy of Agricultural Sciences , B eij ing 100094) Ab stract : A cDNA encoding aminopeptidase N was cloned by degenerated PCR combined with RACE technique . The fulllength of APNHarm was 3 043bp . Open reading frame is 2 856 bp in length , encoding 951 amino acid residues. Its predicted molecular weight and isoelectric point were 108. 3 kD and 5. 29 , respectively . This deduced amino acid sequence shared some common structural features with aminopeptidase N from several moth species , including the consensus zincbinding motif HEXXHX18 E and the GAMEN motif common to gluzincin aminopeptidases. The first 20 amino acid residues at Cend is hydrophobic transmembrane helix. The sequence of APNHarm was deposited in GenBank and the accession number is AY181026 . Key word s : Helicoverp a armigera ; Aminopeptidase N ; Gene cloning 　　苏方金芽孢杆菌 B acillus thuringiensis 能产生具 中肠细胞的修复作用等 。从现有的研究结果来看 , 有杀虫活性的晶体蛋白。晶体蛋白发挥