ASSIGNMENT OF 30 MICROSATELLITE LOCI TO THE LINKAGE MAP OF ARABIDOPSIS
ASSIGNMENT OF 30 MICROSATELLITE LOCI TO THE LINKAGE MAP OF ARABIDOPSIS
复制标题
DOI：
10.1006/geno.1994.1023
复制
发表时间：
1994-01-01
期刊：
影响因子：
4.4
通讯作者：
ECKER, JR
中科院分区：
文献类型：
--
作者：
BELL, CJ;ECKER, JR
30个微卫星位点被分配到拟南芥连锁图谱。通过检索EMBL和GenBank数据库发现了拟南芥DNA中的几个微卫星序列，随后通过聚合酶链反应（PCR）发现了其中一些微卫星序列用于检测不同拟南芥品系之间的多态性。在拟南芥中存在微卫星及其在遗传作图中的应用得到证实后，对标记选择的质粒文库和小插入基因组文库进行了（CA）（n）和（GA）（n）序列的系统筛选。对阳性克隆进行测序，合成重复序列侧翼的PCR引物，并对不同菌株进行PCR以寻找有用的多态性。令人惊讶的是，在18个（CA）（n）重复序列（n > 13）中，只有一个是多态性的。与此相反，30个（GA）（n）重复序列中有25个、3个（AT）（n）重复序列中有2个、4个（A）（n）重复序列中有2个具有多态性。大多数（CA）（n）重复序列是复杂的，具有相邻的短的二核苷酸、三核苷酸或四核苷酸重复序列，而大多数（GA）（n）、（TA）（n）和（A）（n）重复序列是简单的。（CA）（n）重复序列也是PCR分析难处理的，需要广泛优化PCR条件，而其他重复序列类别大多数用单组标准条件扩增。当检测到多态性时，使用一组源自哥伦比亚和兰茨贝格直立株之间杂交的重组近交系对微卫星进行作图。(C)1994年出版社出版。
Thirty microsatellite loci were assigned to the Arabidopsis linkage map. Several microsatellite sequences in Arabidopsis DNA were found by searching the EMBL and GenBank databases, and a number of these were subsequently found to detect polymorphisms between different Arabidopsis strains by the polymerase chain reaction (PCR). After the presence of microsatellites in Arabidopsis and their utility for genetic mapping had been demonstrated, systematic screening for (CA)(n) and (GA)(n) sequences was carried out on marker-selected plasmid libraries and a small-insert genomic library. Positive clones were sequenced, PCR primers flanking the repeats were synthesized, and PCR was carried out on different strains to look for useful polymorphisms. Surprisingly, of 18 (CA)(n) repeats (n > 13), only one was polymorphic. In contrast, 25 of 30 (GA)(n) repeats, 2 of 3 (AT)(n) repeats, and 2 of 4 (A)(n) repeats were polymorphic. The majority of the (CA)(n) repeats were complex, with adjacent short di-, tri-, or tetranucleotide repeats, whereas most of the (GA)(n), (TA)(n), and (A)(n) repeats were simple. The (CA)(n) repeats were also refractory to PCR analysis, requiring extensive optimization of PCR conditions, whereas the other repeat classes were mostly amplified with a single set of standard conditions. When polymorphisms were detected, the microsatellites were mapped using a set of recombinant inbred lines originating from a cross between the strains Columbia and Landsberg erecta. (C) 1994 Academic Press,Inc.