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淡色库蚊抗药性相关基因--NYD-MLC2基因的克隆与分析
引用本文:钱瑾,孙静,孙立新,孙艳,胡小邦,马磊,朱昌亮,QIAN Jin,SUN Jing,SUN Li-xin,SUN Yan,HU Xiao-bang,MA Lei,ZHU Chang-liang.淡色库蚊抗药性相关基因--NYD-MLC2基因的克隆与分析[J].南京医科大学学报,2006,26(7):473-476.
作者姓名:钱瑾  孙静  孙立新  孙艳  胡小邦  马磊  朱昌亮  QIAN Jin  SUN Jing  SUN Li-xin  SUN Yan  HU Xiao-bang  MA Lei  ZHU Chang-liang
作者单位:南京医科大学病原生物学系 江苏南京210029
基金项目:中国科学院资助项目;江苏省高校自然科学基金
摘    要:目的:获取淡色库蚊抗药性相关基因NYD-MLC2的cDNA全长序列并鉴定其在抗性品系和敏感品系中的表达差异。方法:根据抑制性差减杂交(SSH)结合cDNA芯片分离获得的淡色库蚊抗药性相关NYD-MLC2基因片段序列设计引物,采用快速扩增cDNA末端法(rapidamplificationofcDNAends,RACE)扩增NYD-MLC2基因3′、5′端,经对位拼接获得全长序列,并进行生物信息学分析;荧光定量PCR验证此基因在抗性和敏感品系的表达差异。结果:获得淡色库蚊NYD-MLC2cDNA全长序列,开放阅读框为630bp(GenBank/NCBIDQ140391),编码210个氨基酸。蛋白质序列分析显示,NYD-MLC2与冈比亚按蚊(Anophelesgambiae)一个未知功能的蛋白同源性最高,为91%;实时荧光定量PCR结果显示,NYD-MLC2在淡色库蚊溴氰菊酯抗性品系中的表达量是敏感品系的4.08倍。结论:获得淡色库蚊抗药性相关NYD-MLC2基因cDNA全长序列,并证实其在淡色库蚊溴氰菊酯抗性品系中高表达,提示NYD-MLC2与淡色库蚊溴氰菊酯抗药性相关,值得进一步研究。

关 键 词:淡色库蚊  抗药性  NYD-MLC2基因  定量PCR
文章编号:1007-4368(2006)07-0473-04
收稿时间:2006-01-09
修稿时间:2006年1月9日

Cloning and sequence analysis of a novel deltamethrin-resistance associated gene NYD-MLC2 from Culex pipiens pallens
QIAN Jin,SUN Jing,SUN Li-xin,SUN Yan,HU Xiao-bang,MA Lei,ZHU Chang-liang.Cloning and sequence analysis of a novel deltamethrin-resistance associated gene NYD-MLC2 from Culex pipiens pallens[J].Acta Universitatis Medicinalis Nanjing,2006,26(7):473-476.
Authors:QIAN Jin  SUN Jing  SUN Li-xin  SUN Yan  HU Xiao-bang  MA Lei  ZHU Chang-liang
Abstract:Objective: To obtain a full length cDNA of novel deltamethrin-resistance related gene from Culex pipiens pallens and identify differential expression levels between resistant and susceptible strains.Methods:By RACE(rapid amplifyication of cDNA ends),the full length of NYD-MLC2 cDNA of Cx.pipiens pallens was acquired,then sequenced and analyzed by softwares of bioinformatics.The expression levels of NYD-MLC2 in resistant and susceptible strains were detected by real time quantitative PCR.Results: A sequence of 1 002 base pairs including an open reading frame of 630 base pairs was obtained and named as NYD-MLC2(GenBank/NCBI accession number DQ140391).NYD-MLC2 was found to be expressed 4.08 fold higher in the resistant strain than in the susceptible strain.The encoded protein of Cx.pipiens pallens had 210 amino acids with 91% homology of Anopheles gambiae.Conclusion: NYD-MLC2 expressed higher in resistant strain than in susceptible strain of the Cx.pipiens pallens,indicating that NYD-MLC2 may be associated with the deltamethrin resistance of Cx.pipiens pallens.The results are valuable for further study of the relationship between NYD-MLC2 gene and insect resistance.
Keywords:RACE
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