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Cloning and expression of the Erwinia carotovora subsp. carotovora gene encoding the low-molecular-weight bacteriocin carocin S1.
J Bacteriol. 2007 Jan; 189(2):620-6.JB

Abstract

The purpose of this study was to clone the carocin S1 gene and express it in a non-carocin-producing strain of Erwinia carotovora. A mutant, TH22-10, which produced a high-molecular-weight bacteriocin but not a low-molecular-weight bacteriocin, was obtained by Tn5 insertional mutagenesis using H-rif-8-2 (a spontaneous rifampin-resistant mutant of Erwinia carotovora subsp. carotovora 89-H-4). Using thermal asymmetric interlaced PCR, the DNA sequence from the Tn5 insertion site and the DNA sequence of the contiguous 2,280-bp region were determined. Two complete open reading frames (ORF), designated ORF2 and ORF3, were identified within the sequence fragment. ORF2 and ORF3 were identified with the carocin S1 genes, caroS1K (ORF2) and caroS1I (ORF3), which, respectively, encode a killing protein (CaroS1K) and an immunity protein (CaroS1I). These genes were homologous to the pyocin S3 gene and the pyocin AP41 gene. Carocin S1 was expressed in E. carotovora subsp. carotovora Ea1068 and replicated in TH22-10 but could not be expressed in Escherichia coli (JM101) because a consensus sequence resembling an SOS box was absent. A putative sequence similar to the consensus sequence for the E. coli cyclic AMP receptor protein binding site (-312 bp) was found upstream of the start codon. Production of this bacteriocin was also induced by glucose and lactose. The homology search results indicated that the carocin S1 gene (between bp 1078 and bp 1704) was homologous to the pyocin S3 and pyocin AP41 genes in Pseudomonas aeruginosa. These genes encode proteins with nuclease activity (domain 4). This study found that carocin S1 also has nuclease activity.

Authors+Show Affiliations

Department of Chemistry, National Chung-Hsing University, 250 Kuo Kuang Rd., Taichung, Taiwan 402, Republic of China. chuang@nchu.edu.twNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

17071754

Citation

Chuang, Duen-yau, et al. "Cloning and Expression of the Erwinia Carotovora Subsp. Carotovora Gene Encoding the Low-molecular-weight Bacteriocin Carocin S1." Journal of Bacteriology, vol. 189, no. 2, 2007, pp. 620-6.
Chuang DY, Chien YC, Wu HP. Cloning and expression of the Erwinia carotovora subsp. carotovora gene encoding the low-molecular-weight bacteriocin carocin S1. J Bacteriol. 2007;189(2):620-6.
Chuang, D. Y., Chien, Y. C., & Wu, H. P. (2007). Cloning and expression of the Erwinia carotovora subsp. carotovora gene encoding the low-molecular-weight bacteriocin carocin S1. Journal of Bacteriology, 189(2), 620-6.
Chuang DY, Chien YC, Wu HP. Cloning and Expression of the Erwinia Carotovora Subsp. Carotovora Gene Encoding the Low-molecular-weight Bacteriocin Carocin S1. J Bacteriol. 2007;189(2):620-6. PubMed PMID: 17071754.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cloning and expression of the Erwinia carotovora subsp. carotovora gene encoding the low-molecular-weight bacteriocin carocin S1. AU - Chuang,Duen-yau, AU - Chien,Yung-chei, AU - Wu,Huang-Pin, Y1 - 2006/10/27/ PY - 2006/10/31/pubmed PY - 2007/3/3/medline PY - 2006/10/31/entrez SP - 620 EP - 6 JF - Journal of bacteriology JO - J Bacteriol VL - 189 IS - 2 N2 - The purpose of this study was to clone the carocin S1 gene and express it in a non-carocin-producing strain of Erwinia carotovora. A mutant, TH22-10, which produced a high-molecular-weight bacteriocin but not a low-molecular-weight bacteriocin, was obtained by Tn5 insertional mutagenesis using H-rif-8-2 (a spontaneous rifampin-resistant mutant of Erwinia carotovora subsp. carotovora 89-H-4). Using thermal asymmetric interlaced PCR, the DNA sequence from the Tn5 insertion site and the DNA sequence of the contiguous 2,280-bp region were determined. Two complete open reading frames (ORF), designated ORF2 and ORF3, were identified within the sequence fragment. ORF2 and ORF3 were identified with the carocin S1 genes, caroS1K (ORF2) and caroS1I (ORF3), which, respectively, encode a killing protein (CaroS1K) and an immunity protein (CaroS1I). These genes were homologous to the pyocin S3 gene and the pyocin AP41 gene. Carocin S1 was expressed in E. carotovora subsp. carotovora Ea1068 and replicated in TH22-10 but could not be expressed in Escherichia coli (JM101) because a consensus sequence resembling an SOS box was absent. A putative sequence similar to the consensus sequence for the E. coli cyclic AMP receptor protein binding site (-312 bp) was found upstream of the start codon. Production of this bacteriocin was also induced by glucose and lactose. The homology search results indicated that the carocin S1 gene (between bp 1078 and bp 1704) was homologous to the pyocin S3 and pyocin AP41 genes in Pseudomonas aeruginosa. These genes encode proteins with nuclease activity (domain 4). This study found that carocin S1 also has nuclease activity. SN - 0021-9193 UR - https://www.unboundmedicine.com/medline/citation/17071754/Cloning_and_expression_of_the_Erwinia_carotovora_subsp__carotovora_gene_encoding_the_low_molecular_weight_bacteriocin_carocin_S1_ L2 - http://jb.asm.org/cgi/pmidlookup?view=long&pmid=17071754 DB - PRIME DP - Unbound Medicine ER -