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Cloning, purification, and functional characterization of Carocin S2, a ribonuclease bacteriocin produced by Pectobacterium carotovorum.
BMC Microbiol. 2011 May 12; 11:99.BM

Abstract

BACKGROUND

Most isolates of Pectobacterium carotovorum subsp. carotovorum (Pcc) produce bacteriocins. In this study, we have determined that Pcc strain F-rif-18 has a chromosomal gene encoding the low-molecular-weight bacteriocin, Carocin S2, and that this bacteriocin inhibits the growth of a closely related strain. Carocin S2 is inducible by ultraviolet radiation but not by mutagenic agents such as mitomycin C.

RESULTS

A carocin S2-defective mutant, TF1-2, was obtained by Tn5 insertional mutagenesis using F-rif-18. A 5706-bp DNA fragment was detected by Southern blotting, selected from a genomic DNA library, and cloned to the vector, pMS2KI. Two adjacent complete open reading frames within pMS2KI were sequenced, characterized, and identified as caroS2K and caroS2I, which respectively encode the killing protein and immunity protein. Notably, carocin S2 could be expressed not only in the mutant TF1-2 but also in Escherichia coli DH5α after entry of the plasmid pMS2KI. Furthermore, the C-terminal domain of CaroS2K was homologous to the nuclease domains of colicin D and klebicin D. Moreover, SDS-PAGE analysis showed that the relative mass of CaroS2K was 85 kDa and that of CaroS2I was 10 kDa.

CONCLUSION

This study shown that another nuclease type of bacteriocin was found in Pectobacterium carotovorum. This new type of bacteriocin, Carocin S2, has the ribonuclease activity of CaroS2K and the immunity protein activity of CaroS2I.

Authors+Show Affiliations

Department of Chemistry, National Chung-Hsing University, Taichung, 402, Taiwan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21569432

Citation

Chan, Yung-Chieh, et al. "Cloning, Purification, and Functional Characterization of Carocin S2, a Ribonuclease Bacteriocin Produced By Pectobacterium Carotovorum." BMC Microbiology, vol. 11, 2011, p. 99.
Chan YC, Wu JL, Wu HP, et al. Cloning, purification, and functional characterization of Carocin S2, a ribonuclease bacteriocin produced by Pectobacterium carotovorum. BMC Microbiol. 2011;11:99.
Chan, Y. C., Wu, J. L., Wu, H. P., Tzeng, K. C., & Chuang, D. Y. (2011). Cloning, purification, and functional characterization of Carocin S2, a ribonuclease bacteriocin produced by Pectobacterium carotovorum. BMC Microbiology, 11, 99. https://doi.org/10.1186/1471-2180-11-99
Chan YC, et al. Cloning, Purification, and Functional Characterization of Carocin S2, a Ribonuclease Bacteriocin Produced By Pectobacterium Carotovorum. BMC Microbiol. 2011 May 12;11:99. PubMed PMID: 21569432.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Cloning, purification, and functional characterization of Carocin S2, a ribonuclease bacteriocin produced by Pectobacterium carotovorum. AU - Chan,Yung-Chieh, AU - Wu,Jian-Li, AU - Wu,Huang-Pin, AU - Tzeng,Kuo-Ching, AU - Chuang,Duen-Yau, Y1 - 2011/05/12/ PY - 2010/09/21/received PY - 2011/05/12/accepted PY - 2011/5/17/entrez PY - 2011/5/17/pubmed PY - 2011/9/9/medline SP - 99 EP - 99 JF - BMC microbiology JO - BMC Microbiol VL - 11 N2 - BACKGROUND: Most isolates of Pectobacterium carotovorum subsp. carotovorum (Pcc) produce bacteriocins. In this study, we have determined that Pcc strain F-rif-18 has a chromosomal gene encoding the low-molecular-weight bacteriocin, Carocin S2, and that this bacteriocin inhibits the growth of a closely related strain. Carocin S2 is inducible by ultraviolet radiation but not by mutagenic agents such as mitomycin C. RESULTS: A carocin S2-defective mutant, TF1-2, was obtained by Tn5 insertional mutagenesis using F-rif-18. A 5706-bp DNA fragment was detected by Southern blotting, selected from a genomic DNA library, and cloned to the vector, pMS2KI. Two adjacent complete open reading frames within pMS2KI were sequenced, characterized, and identified as caroS2K and caroS2I, which respectively encode the killing protein and immunity protein. Notably, carocin S2 could be expressed not only in the mutant TF1-2 but also in Escherichia coli DH5α after entry of the plasmid pMS2KI. Furthermore, the C-terminal domain of CaroS2K was homologous to the nuclease domains of colicin D and klebicin D. Moreover, SDS-PAGE analysis showed that the relative mass of CaroS2K was 85 kDa and that of CaroS2I was 10 kDa. CONCLUSION: This study shown that another nuclease type of bacteriocin was found in Pectobacterium carotovorum. This new type of bacteriocin, Carocin S2, has the ribonuclease activity of CaroS2K and the immunity protein activity of CaroS2I. SN - 1471-2180 UR - https://www.unboundmedicine.com/medline/citation/21569432/Cloning_purification_and_functional_characterization_of_Carocin_S2_a_ribonuclease_bacteriocin_produced_by_Pectobacterium_carotovorum_ L2 - https://bmcmicrobiol.biomedcentral.com/articles/10.1186/1471-2180-11-99 DB - PRIME DP - Unbound Medicine ER -