摘要
以甜樱桃‘红灯’为试材,应用选择性扩增微卫星(SAM)法分离、克隆了100个SSR序列,其中81个非重复,可用。加上搜索数据库所获得的1个SSR序列,一共82个序列用于特异引物的设计。仅从69个序列的77个基因座设计出特异引物。合成38对特异引物,对其中的36个基因座进行检测。其中19对引物扩增出相应大小的片段,另外8对引物扩增出非预期片段。最后,以27个甜樱桃种质的基因组DNA为模板,从27对可扩增出带的引物中,筛选出多态性引物24对,获得了24个甜樱桃基因座特异性SSR标记。
A total of 100 sequences were isolated and cloned by SAM （ Selectively Amplified Microsatellite） and another one was obtained from the NCBI and EMBL databases. 82 SSR sequences were used to design the special primers at 77 loci from 69 fragments. Thirty-eight pairs of special primers were synthesized, matching with 5＇ anchored degenerate SSR primer, to detect 36 SSR loci. 27 primer pairs amplified clear and robust DNA fragment, of which nineteen pairs of SSR primers amplified the corresponding SSR sequences and eight amplified the unexpected fragments. 24 polymorphic primer pairs were selected from the 27 primer pairs by using the genomic DNA of 27 sweet cherry germplasm, and 24 locus-specific SSR markers were obtained.
出处
《园艺学报》
CAS
CSCD
北大核心
2007年第2期311-316,共6页
Acta Horticulturae Sinica
基金
Supported by the Youth Foundation of Shandong Academy of Agricultural Sciences(2005YQ013)
International Science and Technology Co-operation Projects(2006DFA33130)
by the Chinese National Programs for High Technology and Development(2006AA100108)
关键词
甜樱桃
种质
SSR标记
SAM法
Prunus avium L.
Germplasm
SSR markers
Selectively Amplified Microsatellite
