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棉铃虫性染色体两种分子标记的克隆及序列分析
引用本文:牛宝龙,翁宏飚,何丽华,沈卫锋,孟智启.棉铃虫性染色体两种分子标记的克隆及序列分析[J].昆虫学报,2007,50(7):649-654.
作者姓名:牛宝龙  翁宏飚  何丽华  沈卫锋  孟智启
作者单位:(浙江省农业科学院蚕桑研究所,杭州 310021)
基金项目:国家科学技术部国际科技合作重点项目
摘    要:为了建立棉铃虫Helicoverpa armigera性染色体的特异性分子标记,利用RAPD-PCR技术对雌雄棉铃虫基因组DNA进行筛选,从500种随机引物中筛选到1 条引物(Operon编号为AF-18),可扩增出1条约450 bp 的雌性特异片段。经克隆测序并合成特异引物进行验证,表明该片段为棉铃虫雌性特异分子标记,位于W染色体上。利用家蚕、果蝇等昆虫Kettin基因序列,克隆了棉铃虫的同源基因HaKettin片段,并采用荧光定量PCR技术,以棉铃虫的DH-PBAN基因为参照基因,检测棉铃虫雌雄不同个体间HaKettin基因与DH-PBAN基因的拷贝数之比,结果表明:雄体HaKettinDH-PBAN=1.0,雌体HaKettinDH-PBAN=0.5,据此推断HaKettin基因位于棉铃虫Z染色体上。

关 键 词:棉铃虫  随机扩增多态性DNA  分子标记  性染色体  HaKettin基因  
文章编号:0454-6296(2007)07-0649-06
收稿时间:2006-12-20
修稿时间:2007-04-10

Cloning and sequencing of two molecular markers located on the sex chromosomes of the cotton bollworm, Helicoverpa armigera
NIU Bao-Long,WENG Hong-Biao,HE Li-Hua,SHEN Wei-Feng,MENG Zhi-Qi.Cloning and sequencing of two molecular markers located on the sex chromosomes of the cotton bollworm, Helicoverpa armigera[J].Acta Entomologica Sinica,2007,50(7):649-654.
Authors:NIU Bao-Long  WENG Hong-Biao  HE Li-Hua  SHEN Wei-Feng  MENG Zhi-Qi
Institution:(Sericulture Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China)
Abstract:The individual genome DNA of male and female cotton bollworms (Helicoverpa armigera) was screened with RAPD-PCR technology by 500 random primers. A 450 bp marker amplified from female genome DNA with random primer Operon AF-18 was cloned and sequenced, which was confirmed as a specific molecular marker on the W chromosome. A part of HaKettin gene was cloned according to the sequences of homologues from Bombyx mori and Drosophila, and its copy number in the genome DNA of male and female cotton bollworms was detected with the real-time quantitative PCR technique compared with DH-PBAN. The ratio of copy between HaKettin and DH-PBAN was 1.0 in males and 0.5 in females, showing that the copy number of HaKettin is equal to that of Z chromosome in the genome of cotton bollworm. It is so inferred that HaKettin is located on Z chromosome in the genome of cotton bollworm.
Keywords:Helicoverpa armigera  RAPD  molecular marker  sex chromosome  HaKettin gene
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