ABUNDANCE AND CHARACTERIZATION OF SIMPLE-SEQUENCE REPEATS (SSRS) ISOLATED FROM A SIZE-FRACTIONATED GENOMIC LIBRARY OF BRASSICA-NAPUS L (RAPESEED)
ABUNDANCE AND CHARACTERIZATION OF SIMPLE-SEQUENCE REPEATS (SSRS) ISOLATED FROM A SIZE-FRACTIONATED GENOMIC LIBRARY OF BRASSICA-NAPUS L (RAPESEED)
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DOI：
10.1007/bf00220879
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发表时间：
1995-07-01
影响因子：
5.4
通讯作者：
MCFERSON, JR
中科院分区：
文献类型：
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作者：
KRESOVICH, S;SZEWCMCFADDEN, AK;MCFERSON, JR
对甘蓝型油菜(Brassica napus L.，简称油菜)15 000个克隆的大小分级文库进行了筛选，发现存在GA、CA和GATA简单序列重复序列(SSR)。GA-SSR的丰度分别是CA-SSR和GATA-SSR的四倍和五倍，其出现频率约为每100kb DNA对应一个SSR。对124个阳性克隆进行测序后，设计并评价了7个SSR的引物对。在甘蓝型油菜、甘蓝和油菜的一系列个体中扩增产物，表明这7个SSR在物种间是保守的。两个SSR是多态的。在11份材料中，二核苷酸重复序列B.n.9A扩增出12个片段，四核苷酸重复序列B.n.6A2扩增出两个片段。自动、基于荧光的聚丙烯酰胺凝胶检测已被用于同时增加产量、降低单位成本、提高分析分辨率，并加快SSR分析的数据采集。虽然最初的资金投入和技术能力可能会阻止一些人直接采用我们记录的方法，但SSR分析作为加强遗传资源保护和利用的基因研究工具，值得进一步研究。
A size-fractionated library of Brassica napus L. (rapeseed), composed of 15 000 clones, was screened for the presence of GA-, CA-, and GATA-simple-sequence repeats (SSRs). GA-SSRs were four- and five-fold more abundant than CA- and GATA-SSRs, respectively, and present at a frequency of approximately one SSR for every 100 kb of DNA. Following the sequencing of 124 positive clones, primer pairs were designed and evaluated for seven selected SSRs. Products were amplified in an array of individuals of B. napus, B. oleracea and B. rapa, demonstrating that the seven SSRs were conserved among species. Two SSRs were polymorphic. Among 11 accessions, the dinucleotide (GA)-repeat, B.n.9A, yielded 12 fragments, while the tetranucleotide-repeat (GATA), B.n.6A2, revealed two fragments. Automated, fluorescence-based detection of polyacrylamide gels has been employed to simultaneously increase throughput, reduce unit cost, improve analytical resolution, and expedite data acquisition of SSR analysis. Though initial financial investment and technical capabilities may prevent some from directly employing our documented approach, SSR analysis warrants further investigation as a tool in genetic studies for enhancing both the conservation and utilization of genetic resources.