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Complete nucleotide sequence of pSMB 74, a plasmid encoding the production of pediocin AcH in Pediococcus
A Motlagh1, M Bukhtiyarova, B Ray
1Department of Animal Science, University of Wyoming, Laramie 82071.
Abstract:
Several Pediococcus acidilactici strains produce a plasmid-encoded bacteriocin, pediocin AcH. Previous studies have shown that this plasmid, designated as pSMB 74, encodes genes associated with the production of prepediocin, its post-translation processing to pediocin AcH, transmembrane translocation of these molecules, and immunity of producer cells against pediocin AcH. We report here the complete nucleotide sequence of pSMB 74. The plasmid has a total of 8877 bp. Four genes have been located on pSMB 74. The genes are arranged in a gene cluster of 3500 bp and share a common promoter and rho-independent stem-loop terminator. The four genes, each with independent ribosome binding sites (rbs), initiation and termination codons and spacer sequences in between, were designated as pap A, pap B, pap C and pap D and encode respectively for proteins of 62, 112, 174 and 724 amino acids. The results of this study can be useful either to introduce a suitable marker at a unique restriction site in pSMB 74 and use it as a vector or to clone the pap gene cluster in a suitable plasmid and transform desirable strains for pediocin AcH production. The gene sequence has been submitted to Gene Bank (Acc. No. U02482).
Insights
The complete nucleotide sequence of the Pediococcus acidilactici plasmid pSMB 74 was determined. This plasmid encodes four genes (pap A-D) essential for pediocin AcH bacteriocin production and immunity.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Pediococcus acidilactici produces pediocin AcH, a bacteriocin encoded by the plasmid pSMB 74.
- The plasmid contains genes for prepediocin production, processing, translocation, and cell immunity.
Purpose of the Study:
- To determine the complete nucleotide sequence of the pSMB 74 plasmid.
- To identify and characterize the genes involved in pediocin AcH production.
Main Methods:
- Plasmid DNA sequencing.
- Bioinformatic analysis to identify genes and regulatory elements.
Main Results:
- The complete nucleotide sequence of pSMB 74 (8877 bp) was determined.
- Four genes (pap A, pap B, pap C, pap D) were identified within a 3500 bp gene cluster.
- These genes encode proteins of varying sizes and are regulated by a common promoter and terminator.
Conclusions:
- The sequence data provides a foundation for using pSMB 74 as a cloning vector or for transferring the pediocin AcH gene cluster to other strains.
- Understanding the genetic organization facilitates biotechnological applications for bacteriocin production.