• Genomics & Informatics
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• 제1권1호
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• pp.55-60
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• 2003

The nucleotide sequence of pZMO1, a small cryptic plasmid of Zymomonas mobilis ATCC10988 was determined. Analysis of 1,680 bp of sequence revealed $69\%$ identity with Shigella sonnei plasmid, pKYM and $61\%$ identity with Nostoc sp. ss DNA replicating plasmid. Analysis of a deduced amino acid sequence of an orf of pZMO1 revealed $75\%$ identity and $90\%$ similarity with the repA gene of Synechocystis sp. plasmid pCA2.4. The upstream region of the repA gene of pZMO1 possesses six directed repeat sequences and two inverted repeat sequences at downstream of the IR consensus sequence of nick region of rolling circle replication (RCR) plasmid. A typical terminator hairpin structure was found at the downstream region of repA gene. Degradation of single-stranded plasmid DNA by S1 nuclease was detected by Southern hybridization. It suggests that pZMO1 replicates by a rolling circle mechanism in Z. mobilis ATCC10988 cells.

• 식물병연구
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• 제15권2호
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• pp.77-82
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• 2009

최근 혹병이 심하게 발생한 충북지방의 포도나무의 뿌리로부터 선택배지를 이용하여 Agrobacterium속 세균을 분리하고 동정하였다. 분리균의 지방산 분석을 통한 MIDI에 의한 동정, 16S rDNA 염기서열, 생화학적 특성, 종 특이적 primer을 이용한 PCR 결과로 13개 분리균은 모두 A. tumefaciens로 동정 되었으며, 포도나무 혹병균인 A. vitis는 분리되지 않았다. 모든 분리균은 토마토와 포도나무의 줄기와 뿌리에 병원성을 나타내지 않았다. Rep-PCR 결과 분리균은 A. tumefaciens와 일부 유사성이 있지만 전체적으로 병원균인 A. tumefaciens와 A. vitis와는 유사성이 낮았다. 이상의 결과는 충북 일부 지방의 MBA와 거봉에서 발생한 혹병을 일으키는 병원균은 토양이나 뿌리로부터 전반되지 않고 묘목을 통해서 전반되었을 가능성을 시사하고 있다.

• Journal of Microbiology and Biotechnology
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• 제33권2호
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• pp.211-218
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• 2023

A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Em^r), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.