Tourmente S, Deragon J M, Lafleuriel J, Tutois S, Pélissier T, Cuvillier C, Espagnol M C, Picard G
GDR 977 Biomove CNRS, Université Blaise Pascal, Aubiere, France.
Nucleic Acids Res. 1994 Aug 25;22(16):3317-21. doi: 10.1093/nar/22.16.3317.
A strategy based on random PCR amplification was used to isolate new repetitive elements of Arabidopsis thaliana. One of the random PCR product analyzed by this approach contained a tandem repetitive minisatellite sequence composed of 33 bp repeated units. The genomic locus corresponding to this PCR product was isolated by screening a lambda genomic library. New related loci were also isolated from the genomic library by screening with a 14 mer oligonucleotide representing a region conserved among the different repeated units. Alignment of the consensus sequence for each minisatellite locus allowed the definition of an Arabidopsis thaliana core sequence that shows strong sequence similarities with the human core sequence and with the generalized recombination signal Chi of Escherichia coli. The minisatellites were tested for their ability to detect polymorphism, and their chromosomal position was established.
一种基于随机PCR扩增的策略被用于分离拟南芥新的重复元件。通过这种方法分析的一个随机PCR产物包含一个由33 bp重复单元组成的串联重复微卫星序列。通过筛选λ基因组文库分离出与该PCR产物对应的基因组位点。还通过用代表不同重复单元间保守区域的14聚体寡核苷酸筛选基因组文库，从基因组文库中分离出了新的相关位点。对每个微卫星位点的共有序列进行比对，确定了一个拟南芥核心序列，该序列与人类核心序列以及大肠杆菌的通用重组信号Chi具有很强的序列相似性。测试了微卫星检测多态性的能力，并确定了它们在染色体上的位置。
