椰子织蛾CSP基因的克隆与序列分析
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摘要
利用rapid-amplification of cDNA ends(RACE)技术,克隆获得了1条椰子织蛾(Opisina aremosella)的全长化感蛋白基因序列。该基因全长为561 bp,开放阅读框全长为393 bp,3′和5′端非编码序列分别为48、120 bp,编码130个氨基酸,所编码蛋白质的理论分子量为14.8 ku,等电点为5.85。同源性比对分析发现,椰子织蛾化感蛋白基因氨基酸序列有4个保守半胱氨酸位点且与其他昆虫化感蛋白基因在氨基酸的水平上相似性不高。除与棉红铃虫(Pectinophora gossypiella)相似度达71%外,与其他昆虫相比,均低于10%。SignalP预测显示,椰子织蛾化感蛋白基因所编码的前20个氨基酸为信号肽序列(MAMKYLIVLSCVLAAAIVAG)。系统进化树分析结果表明,椰子织蛾化感蛋白与稻纵卷叶螟(Cnaphalocrocis medinalis)、亚洲玉米螟(Ostrinia furnacalis)和家蚕(Bombyx mori)化感蛋白1、2、3、4聚为同一族,与家蚕化感蛋白1、2、3、4的进化程度最近。这为深入研究昆虫化学感受蛋白、揭开昆虫与环境化学信息联系的规律奠定理论基础。
引文
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[3]Pelosi P,Zhou J J,Ban L P,et al. Soluble proteins in insect chemical communication[J]. Cellular and Molecular Life Sciences,2006,63(14):1658-1676.
[4]王思豹,张帅,雒珺瑜,等. 大草蛉化学感受蛋白基因CpalCSP3组织表达谱及气味结合特性分析[J]. 棉花学报,2015,27(3):260-267.
[5]Natori S,Kubo T,Nomura A,et al. Purification and localization of p10,a novel protein that increases in nymphal regenerating legs of Periplaneta americana(American cockroach).[J]. International Journal of Developmental Biology,1992,36(3):391-8.
[6]Kitabayashi A N,Arai T,Kubo T,et al. Molecular cloning of cDNA for p10,a novel protein that increases in the regenerating legs of Periplaneta americana (American cockroach)[J]. Insect Biochemistry and Molecular Biology,1998,28(10):785-790.
[7]柳晓磊,翁群芳,胡美英,等. 昆虫化学感受蛋白基因的进化分析[J]. 华中农业大学学报,2011,2(2):177-181.
[8]阎伟,吕宝乾,李洪,等. 椰子织蛾传入中国及其海南省的风险性分析[J]. 生物安全学报,2013,22(3):163-168.
[9]李洪,刘丽,阎伟. 新入侵害虫椰子织蛾的发生及防治[J]. 中国森林病虫,2015,4(4):10-13.
[10]陆永跃,王敏. 椰子织蛾的形态特征识别[J]. 环境昆虫学报,2013,35(6):838-842.
[11]刘向蕊,吕宝乾,金启安,等. 5种杀虫剂对入侵害虫椰子织蛾的室内毒力测定[J]. 生物安全学报,2014,23(1):13-17.
[12]Thompson J D,Gibson T J,Plewniak F,et al. The CLUSTAL_X Windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Research,1997,25(24):4876-4882.
[13]Dyrl?v B J,Nielsen H,von Heijne G,et al. Improved prediction of signal peptides:SignalP 3.0[J]. Journal of Molecular Biology,2004,340(4):783-795.
[14]Hulo N,Bairoch A,Bulliard V,et al. The 20 years of PROSITE[J]. Nucleic Acids Research,2008,36:D245.
[15]Tamura K,Dudley J,Nei M,et al. MEGA4:molecular evolutionary genetics analysis (MEGA) software version 4.0[J]. Molecular Biology and Evolution,2007,24(8):1596-1599.
[16]Vieira F G,Rozas J . Comparative genomics of the odorant-binding and chemosensory protein gene families across the arthropoda:origin and evolutionary history of the chemosensory system[J]. Genome Biology and Evolution,2011,3:476-490.
[17]刘金香,钟国华,谢建军,等. 昆虫化学感受蛋白研究进展[J]. 昆虫学报,2005,48(3):418-426.白羽,丁海麦,石松利. 濒危植物蒙古扁桃SRAP反应体系优化[J]. 江苏农业科学,2019,47(3):46-48.