黑麦重复家族成员pAW161(348 bp)的DNA序列分析

黑麦亚端部特异序列pAW161是黑麦350~480 bp重复家族的一个成员。通过pAW161设计的特异引物获得的PCR扩增片段,已成功用于小麦外源转移材料的鉴定。在本研究中,用该引物从供试的分属4个种共7份黑麦材料中都扩增出一条清楚明亮的特异带纹,分别对扩增片段回收、克隆、测序。结果表明,它们的长度都为348bp,该序列简称为pAW161(348 bp)。不同黑麦比较发现,该序列高度保守,同源性达到99.39%,可作为黑麦的亚端部特异序列pAW161的分子标记。该序列存在着6个SNP位点,都位于此序列的3’端的156 bp序列中,而5’端的192 bp的序列无变异;SNP变化在供试材料中不存在物种特异性。从结构上看,pAW161(348 bp)是重复家族pSc200的380 bp基本重复单元的两个头-尾串联重复序列的中间一段。

DNA Sequence Analysis of Rye (Secale cereale L. ) Repeated Family pAW161 (348 bp)

pAw161 was a member of rye(Secale cereale L.) subtelomeric repeated sequence 350～480-family.Specific PCR primer designed for pAW161 has been successfully used for detecting introgression of alien chromatin in wheat.By using the primer,only one sharp band was respectively amplified in 7 rye materials in this study.The result from DNA sequencing indicates that the PCR band is of a length of 348 bp.This sequence is designated as pAW161(348 bp),which is highly conserved among the 7 ryes with a similarity of 99.39%.It can be used as a specific marker for pAW161.Six SNP sites are found in this sequence.It is very interesting that the six SNP sites are only distributed on the 156 bp of the 3' sequence end,while the remainder 192 bp of 5' sequence is completely conserved.The variation of SNP sites is not species-specific among the materials in this study.BLAST analysis indicates that pAW161(348 bp) is the middle fragment of the pSc200 repeated family,which is organized by head-to-tail with two 380 bp monomer.