基于RAPD的SCAR分子标记技术鉴定虎纹蛙及引进种

为了对引进种泰国虎纹蛙加强种质监测,本文对泰国虎纹蛙和中国虎纹蛙进行了SCAR标记研究。在对2种蛙随机扩增多态DNA(RAPD)标记研究的基础上,选用随机引物OPP5和OPZ19对2种蛙进行扩增,将2个特异性扩增片段OPP5-471bp和OPZ19-512bp回收、克隆和测序,并根据序列信息设计两对SCAR引物SP5和SZ19。利用两对SCAR引物对2种蛙基因组DNA扩增的结果显示:SP5显示了其特异性,在对中国虎纹蛙的扩增中出现了单一SP5-388bp特异片段,而在泰国虎纹蛙中没有扩增产物;SZ19在对2种虎纹蛙的扩增中出均出现了单一条带,但片段大小不同,在中国虎纹蛙中为SZ19-456bp,在泰国虎纹蛙中为SZ19-497bp。3个分子标记都可作为基于RAPD成功转化后的SCAR分子标记对2种虎纹蛙进行鉴定。

IDENTIFICATION OF INTRODUCED RANA TIGRINA SPECIES BY SCAR MARKERS BASED ON RAPD

To strengthen the monitor of Rana tigrina cantor,identification of Rana tigrina cantor and R.tigrina rugulosa were studied with the SCAR(Sequencecharacterized amplified regions) technique based on RAPD(Random Amplified Polymorphic DNA).Two random Primers,OPP5 and OPZ19,were selected for amplification in R.tigrina cantor and R.tigrina rugulosa.Specific amplified DNA fragments,OPP5-471bp and OPZ19-512bp,were found,cloned and sequenced.Two SCAR primers,SP5 and SZ19,were designed according to their sequence information.The results showed: 1.there was a specific fragment SP5-388bp in R.tigrina rugulosa and no PCR amplification in R.tigrina cantor. which proves that the SP5-388bp was a species-specific SCAR marker of R.tigrina rugulosa.2.SZ19-456bp and SZ19-497 were obtained in the amplification of R.tigrina rugulosa and R.tigrina cantor,respectively,by SCAR primer SZ19.Three effective SCAR markers were obtained by transformation on the basis of RAPD in this study.