从植物致病土壤杆菌菌株中进行Ti和Ri质粒的PCR检测。
PCR detection of Ti and Ri plasmids from phytopathogenic Agrobacterium strains.
作者信息
Sawada H, Ieki H, Matsuda I
机构信息
National Institute of Agro-Environmental Sciences, Ibaraki, Japan.
出版信息
Appl Environ Microbiol. 1995 Feb;61(2):828-31. doi: 10.1128/aem.61.2.828-831.1995.
Abstract
A universal primer set (VCF/VCR) for PCR analysis based on the sequences of the virC operon located on Ti and Ri plasmids was designed to detect these plasmids from phytopathogenic Agrobacterium strains. With the VCF (sequence, 5'-ATCATTTGTAGCGACT-3') and VCR (sequence, 5'-AGCTCAAACCTGCTTC-3') primer set, DNA fragments of 730 bp in length were amplified from cell lysates of 10 rhizogenic and 65 tumorigenic agrobacteria. DNA sequencing and Southern hybridization analysis confirmed that the amplified fragments corresponded to the target region. The PCR method is considered convenient for routine determination of the potential pathogenicity of Agrobacterium strains.
摘要
基于Ti和Ri质粒上virC操纵子序列设计了用于PCR分析的通用引物组（VCF/VCR），以从植物病原性农杆菌菌株中检测这些质粒。使用VCF（序列：5'-ATCATTTGTAGCGACT-3'）和VCR（序列：5'-AGCTCAAACCTGCTTC-3'）引物组，从10株发根农杆菌和65株致瘤农杆菌的细胞裂解物中扩增出长度为730 bp的DNA片段。DNA测序和Southern杂交分析证实扩增片段对应于目标区域。该PCR方法被认为便于常规测定农杆菌菌株的潜在致病性。
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