云南切梢小蠹细胞色素P450基因的克隆与分析  

Cloning and sequence analysis of cytochrome P450 gene from Tomicus yunnanensis

作　　者：季梅[1] 泽桑梓[1] 

机构地区：[1]云南省林业科学院森林保护研究所,云南昆明650201

出　　处：《广东农业科学》2012年第23期146-148,共3页Guangdong Agricultural Sciences

基　　金：国家林业局林业公益性行业科研专项(201004067)

摘　　要：利用RACE技术，从云南切梢小蠹Tomicus yunnanensis中克隆获得了一个细胞色素P450基因cDNA序列。该基因简写为TyunP450，其序列长1524bp，开放性阅读框为1269bp，编码422个氨基酸，预测蛋白分子量为48．50ku，pI为8．94。通过Motif Scan分析发现，TyunP450氨基酸序列中潜在1个酰胺化位点（XGR265-268）、4个N-糖基化位点（NLSV33—36、NDSV87-90、NGST97—100和NASK372—375）、4个酪蛋白激酶Ⅱ磷酸位点（SVTE176—179、TQRD187—190、SDED217—220和TIDE420—423）、5个N-豆蔻酰化位点（GNVFGF2—7、GNVYND40—45、GGQNAA163—168、GGILSD213—218和GTNIAI340—345）、8个蛋白质激酶C磷酸化位点（SLK77-79、STK99—101、TQR187—189、SLK210—212、SLR288—289、SFIK417—419、SLK431-433和TLR443-445）、1个P450细胞色素血红素铁配体的半胱氨酸位点（FSSGPRDCLG384—393），204～316氨基酸区域具有典型的细胞色素P450结合位点结构，317-441氨基酸区域具有典型的细胞色素P450结合位点结构，9～28氨基酸区域具有DUF321蛋白的未知函数结构域，1-441氨基酸区域具有典型的细胞色素P450结合位点结构。TyunP450与埃及伊蚊Aedesaegypt、家蚕Bombyxmori、丽蝇蛹集金小蜂Nasoniavitripennis、赤拟谷盗Tribolium castarteum和豌豆蚜Acyrthosiphon pisum的P450氨基酸序列相似性分别为19％、20％、21％、23％和15％。In this paper, a cDNA clone encoding cytochrome p450 was firstly isolated from Tomicus yunnanensis using rapid amplification of eDNA ends （RACE） strategy. This gene named TyunP450 sequence was 1 524 bp in full-length. Its open reading frame was 1 269 bp, encoding a protein of 422 amino acid with predicted molecular mass of 48.5 ku and pI of 8.94. Motif Scan analysis indicated that TyunP450 amino acid sequence potentially contained 1 amidation site （XGR265-268）, 4 N glycosylation sites （ NLSV33- 36, NDSV87-90, NGST97-100 and NASK372-375）, 4 casein kinase II phosphorylation sites （SVTE176-179, TQRD187-190, SDED217- 220amt TIDE420-423）, 5 N myristoylation sites （GNVFGF2-7, GNVYND40-45, GGQNAA163-168, GGILSD213-218 and GTNIAI340- 345）, 8 protein kinase C phosphorylation sites （ SLK77-79, STK99-101, TQR187-189, SLK210-212, SLR288-289, SHK417-419, SLK431-433 and TLR443-445）, and 1 P450 cytoehromes heine iron ligand cysteine sites （FSSGPRDCLG384-393）. 204-316 amino acid region has typical eytochrome P450 binding site structure. 317-441 amino acid region had typical cytochrome P450 binding site structure. 9~28 amino acid region was of DUF321 protein of unknown function domain. And 1-441 amino acid region had typical cytochrome P450 binding site structure. The P450 similarities of Tomicus .yunnanensis to that of Aedes aegypti, Bombyx mori, Nasonia vitripennis, Tribolium castaneum, Acyr thosiphon pisum were 19%, 20%, 21%, 23% and 15%, respectively.

关 键 词：云南切梢小蠹 P450 基因克隆 序列分析 

分 类 号：S718.7[农业科学—林学]