与鲤鱼抗寒性状相关的RAPD分子标记的筛选及其克隆  

Screening and cloning of RAPD marker interrelated to cold tolerance in common carp

作　　者：梁利群[1] 高俊生[2] 李绍戊[2] 孙效文[2] 雷清泉[1] 

机构地区：[1]哈尔滨理工大学,黑龙江哈尔滨150080 [2]中国水产科学研究院黑龙江水产研究所

出　　处：《中国水产科学》2006年第3期360-364,共5页Journal of Fishery Sciences of China

基　　金：国家"973"基础研究项目资助(2004CB117404)

摘　　要：以黑龙江鲤(Cyprinus carpio haematopterusTemminck et Schlegel)、荷包红鲤抗寒品系(Cyprinus carpiovar.wana-nensis)、荷包红鲤(Cyprinus carpiovar.wananensis)、柏氏鲤(Cyprinus pellegniniTchang)以及荷包红鲤抗寒品系和柏氏鲤的杂交F2的DNA样品为材料,用200个随机引物对其进行PCR扩增,获得了1个与鲤鱼抗寒性状相关的分子标记RAG20,并在F2代分离个体中得到验证。至此,本实验室共获得10个与鲤鱼抗寒性状相关的分子标记,更加证明抗寒性状是数量性状(QTL)受微效多基因控制。此外,将其中4个标记从琼脂糖凝胶上进行回收,并与pMD18-T vector连接,将其转入感受态大肠杆菌E.coliDH 5α,对克隆片段进行序列分析,旨为根据每一序列重新设计引物,将RAPD标记转化为稳定的SCAR标记。RAPD-PCR was performed with 200 random primers on DNA samples of Heilongjiang wild carp （Cyprinus carpio haematopterus Temminck et Schlegel ）, frigid-resistance strain of Red purse carp （Cyprinus carpio var. wananensis ）, Red purse carp （Cyprlnus carpio vat. wananensis ）, Boshi carp （ Cyprinus pellegnini Tchang）, cross F2 of frigid-resistance strain of Red purse and Boshi carp, respectively. RAG20 was identified to be a molecular marker which associated with cold tolerance traits of common carp （cyprinus carpio）. Examination to F2 segregated individuals verified that this molecular marker was reliable. Up to now, 10 molecular markers associated with cold tolerance of common carp were obtained, and the marker 5N1451c interrelated with cold tolerance Cyprinus （ Cyprinus ） pellegrini Tchang has been mapped on linkage 5. It further demonstrates that cold tolerance trait is a quantitative trait loci （QTL）, which is controlled by small multi-genes. In addition, specific fragments from four primers above were extracted from the agarose and purified respectively. The purified products were then ligated to pMD 18-T vector which were transformed to competent E. coli DH 5α for sequencing. The sequencing of three specific fragmens has been finished. The length of the amplified fragments was 1 021 bp, 422 bp, 1 043 bp respectively. The parpose of this study is to redesign primers upon the sequences and convert RAPD markers into the stable SCAR markers finally.

关 键 词：鲤鱼 抗寒 SCAR RAPD 分子标记 

分 类 号：Q78[生物学—分子生物学]