کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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2090635 | 1081512 | 2009 | 9 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Characterization of pNC1, a small and mobilizable plasmid for use in genetic manipulation of Desulfovibrio africanus Characterization of pNC1, a small and mobilizable plasmid for use in genetic manipulation of Desulfovibrio africanus](/preview/png/2090635.png)
To develop a vector system that facilitates genetic manipulation in Desulfovibrio species, we screened native sulfate-reducing bacteria for small plasmids. A self-replicating plasmid was discovered in Desulfovibrio africanus SR-1. Sequence analysis of this 8568-bp plasmid (pNC1) revealed a G+C content of 47.2% and nine open reading frames. This plasmid has a copy number of six. Compatible hosts include D. africanus and Pseudomonas aeruginosa PA14. Genetic characterization of pNC1 revealed that 53.6% of the plasmid contains genes associated with replication, mobilization, and partitioning. The 1123-bp replicon is composed of a rep gene and four 22-bp iterons. The mobilization operon is composed of three genes with a putative 144-bp oriT. The partitioning operon is composed of parA and parB with a downstream parS. We report the construction of a small pNC1-based cloning vector which transforms D. africanus at high frequencies (~ 1.5 × 103 CFU/μg DNA), is mobilizable at high transfer frequency (4.8 × 10− 4 transconjugants/donor), and is stably maintained under non-selective pressure. This study provides a potential host–vector system for Desulfovibrio gene functional analyses.
Journal: Journal of Microbiological Methods - Volume 79, Issue 1, October 2009, Pages 23–31