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利用基因组自引物PCR从野豌豆属中分离出两个新的串联重复序列家族。

Two new families of tandem repeats isolated from genus Vicia using genomic self-priming PCR.

作者信息

Macas J, Pozárková D, Navrátilová A, Nouzová M, Neumann P

机构信息

Institute of Plant Molecular Biology, Ceské Budejovice, Czech Republic.

出版信息

Mol Gen Genet. 2000 Jun;263(5):741-51. doi: 10.1007/s004380000245.

Abstract

A modified genomic self-priming technique was used for rapid isolation of tandem repeats from several Vicia species. Based on homologies of their nucleotide sequences the newly isolated clones were assigned to two repeat families named VicTR-A and VicTR-B. Both families are rich in AT (74%) and are organized as long blocks of tandemly repeated units. The VicTR-A repeats are characterized by a monomer size of 69 bp, whereas the VicTR-B repeat monomer is about 38 bp long, and the two families do not share significant sequence homology. VicTR sequences show different degrees of amplification (up to 10(6)-10(7) copies/haploid genome) in individual Vicia species and are not amplified in other legumes. The abundances of these repeats do not correlate with genome sizes but are similar in species that belong to the same taxonomic section within the genus Vicia. Primed in situ (PRINS) labeling of metaphase chromosomes of V. pannonica revealed that VicTR-A sequences are located predominantly in the telomeric regions of the short arms of all chromosomes. In contrast, labeling of VicTR-B repeats in V. sativa resulted in mainly intercalary bands of various intensities and only weak telomeric signals.

摘要

一种改良的基因组自引发技术被用于从几种蚕豆属物种中快速分离串联重复序列。基于其核苷酸序列的同源性,新分离的克隆被归为两个重复家族,分别命名为VicTR - A和VicTR - B。这两个家族都富含AT(74%),并由串联重复单元的长片段组成。VicTR - A重复序列的特征是单体大小为69 bp,而VicTR - B重复单体约为38 bp长,且这两个家族没有显著的序列同源性。VicTR序列在单个蚕豆属物种中表现出不同程度的扩增(高达10^6 - 10^7个拷贝/单倍体基因组),而在其他豆科植物中不扩增。这些重复序列的丰度与基因组大小无关,但在蚕豆属内属于同一分类组的物种中相似。对潘诺尼亚蚕豆中期染色体进行原位引物标记(PRINS)显示,VicTR - A序列主要位于所有染色体短臂的端粒区域。相反,对苜蓿蚕豆中VicTR - B重复序列的标记主要产生各种强度的居间带,仅产生微弱的端粒信号。

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