利用反向PCR鉴定沼泽红假单胞菌中野生质粒  

Identification of native plasmid in a Rhodopseudomonas palustris strain by reverse PCR

作　　者：宋智勇[1] 曲媛媛[1] 周集体[1] 

机构地区：[1]大连理工大学环境与生命学院,辽宁大连116024

出　　处：《大连理工大学学报》2009年第1期33-37,共5页Journal of Dalian University of Technology

摘　　要：为准确研究沼泽红假单胞菌的野生质粒及其携带的基因信息，在常规碱裂解法基础上，对沼泽红假单胞菌Rhodopseudomonas palustrisASl．2352中野生质粒提取方法进行了优化，优化方法具有易操作、耗时少、纯度高等特点．所得质粒DNA纯度和质量均满足进一步分子操作的需要．同时，利用反向PCR技术鉴定该野生质粒，并结合生物信息学方法对其进行分析。比传统的质粒丢失法验证和考察质粒更为省时、可靠和直观．证明菌株携带4个野生质粒，其中2．3kb的质粒序列与Escherichiacoli pEC157 Rom-like proteingene有97．8％的同源性．推测1885～1887碱基为复制起点．In order to study Rhodopseudornonas palustris in detail, the plasmid extraction of Rhodopseudomonas palustris AS1. 2352 was optimized, which was easier and took shorter time, and the quality and quantity of the extracted DNA sample were enough for the further molecular procedures. The native plasmids of this strain were identified by reverse PCR combined with bioinformatics. This method is easier and more reliable than traditional ones. There are four native plasmids in this strain, one of which is 2.3 kb in length and shares 97.8% homology with Escherichia coli plasmid pEC157 Rom-like protein gene, and base 1885-1887 is the origin of replication.

关 键 词：反向PCR 野生质粒 沼泽红假单胞菌 质粒提取 

分 类 号：X172[环境科学与工程—环境科学]