Development and characterization of microsatellite markers from chromosome 1-specific DNA libraries of Vicia faba
Development and characterization of microsatellite markers from chromosome 1-specific DNA libraries of Vicia faba
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DOI：
10.1023/a:1016253214182
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发表时间：
2002-01-01
影响因子：
1.5
通讯作者：
Macas, J
中科院分区：
文献类型：
--
作者：
Pozárková, D;Koblizková, A;Macas, J
一种从蚕豆(Vicia Faba L.)所选区域定向检索微卫星标记的综合方法已经开发出来。基因组。该程序基于先进的物理和遗传作图技术，包括以下步骤：1)中期染色体的流动分选，2)构建富含微卫星的染色体特异DNA文库，3)从文库中分离多态的微卫星序列，4)用纯化的个别染色体类型的部分利用聚合酶链式反应检测微卫星的染色体特异性，以及5)将染色体特异标记整合到遗传图谱中。测试了几种分离微卫星克隆的策略，包括用单一或混合探针直接筛选非浓缩文库，以及在一轮或两轮浓缩后筛选文库。最后，通过从最大的田间菜豆染色体(No.1)的选定部分检索新的标记，证明了该方法的有效性。
An integrated approach has been developed for targeted retrieval of microsatellite markers from selected regions of the field bean (Vicia faba L.) genome. The procedure is based on a combination of advanced physical and genetic mapping techniques and includes the following steps: 1) flow-sorting of metaphase chromosomes, 2) construction of microsatellite-enriched chromosome-specific DNA libraries, 3) isolation of polymorphic microsatellite sequences from the libraries, 4) testing chromosome specificity of the microsatellites using polymerase chain reaction with purified fractions of individual chromosome types, and 5) integration of chromosome-specific markers into a genetic map. Several strategies for isolation of microsatellite clones were tested, including direct screening of non-enriched libraries with single or mixed probes and screening of the libraries after one or two rounds of enrichment. Finally, the usefulness of this approach was demonstrated by the retrieval of novel markers from a selected portion of the largest field bean chromosome (No. 1).