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Identification of nucleotide sequence involved in Weissellicin L production.

Chen YS, Lee YS, Wu HC, Chiang CM, Pan SF, Leong KH - Springerplus (2014)

Bottom Line: The obtained results indicated that the mature bacteriocin consists of 29 amino acid residues with a molecular weight of approximately 3205.64 Da.PCR presented a single product and was useful to detect weissellicin L structural gene.A rapid method to detect weissellicin L structural gene was also reported in this study.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Ming Chuan University, No. 5, De-Ming Rd., Gui-Shan Township, Taoyuan County, 333 Taiwan.

ABSTRACT

Background: Weissellicin L, a novel bacteriocin produced by Weissella hellenica 4-7, was previously characterized but its full amino acid sequence remain unknown. The draft genome sequencing analysis of Weissella hellenica 4-7 was performed and the open reading frame (ORF) encoding the weissellicin L was identified and clarified.

Findings: The obtained results indicated that the mature bacteriocin consists of 29 amino acid residues with a molecular weight of approximately 3205.64 Da. A conserved processing site of two glycine residues in positions -1 and -2 was observed in the leader peptides. The possibility that bacteriocin secretion depended on ATP-binding cassette (ABC) transporter was therefore suggested. Furthermore, primers were designed from 5' and 3' flanking sequences of the weissellicin L structural gene. PCR presented a single product and was useful to detect weissellicin L structural gene.

Conclusions: To our knowledge, this is the first report describing the full amino acid sequence of Weissellicin L. A rapid method to detect weissellicin L structural gene was also reported in this study.

No MeSH data available.


Ethidium bromide stained 2% agarose gel of PCR products using weissellicin L gene specific primers. Lane M, a 100-bp DNA ladder marker; Lane 1, W. hellenica 4–7; Lane 2, W. hellenica BCRC 80264T; Lane 3, W. hellenica 203.
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Fig2: Ethidium bromide stained 2% agarose gel of PCR products using weissellicin L gene specific primers. Lane M, a 100-bp DNA ladder marker; Lane 1, W. hellenica 4–7; Lane 2, W. hellenica BCRC 80264T; Lane 3, W. hellenica 203.

Mentions: Primers specific for the weissellicin L gene were designed to perform PCR amplification. A single 457-bp fragment was amplified from the genomic DNA of W. hellenica 4–7 (Figure 2). However, no amplified PCR product was observed from W. hellenica strains BCRC 80264T and 203 (Figure 2). PCR product from W. hellenica 4–7 was sequenced and the nucleotide sequences encoding the target gene was confirmed to be identical as reported (Figure 1). The results suggested that PCR amplification with the weissellicin L-specific primers could be useful to select potential weissellicin L-producing strain from multitudinous W. hellenica strains.Figure 2


Identification of nucleotide sequence involved in Weissellicin L production.

Chen YS, Lee YS, Wu HC, Chiang CM, Pan SF, Leong KH - Springerplus (2014)

Ethidium bromide stained 2% agarose gel of PCR products using weissellicin L gene specific primers. Lane M, a 100-bp DNA ladder marker; Lane 1, W. hellenica 4–7; Lane 2, W. hellenica BCRC 80264T; Lane 3, W. hellenica 203.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4209004&req=5

Fig2: Ethidium bromide stained 2% agarose gel of PCR products using weissellicin L gene specific primers. Lane M, a 100-bp DNA ladder marker; Lane 1, W. hellenica 4–7; Lane 2, W. hellenica BCRC 80264T; Lane 3, W. hellenica 203.
Mentions: Primers specific for the weissellicin L gene were designed to perform PCR amplification. A single 457-bp fragment was amplified from the genomic DNA of W. hellenica 4–7 (Figure 2). However, no amplified PCR product was observed from W. hellenica strains BCRC 80264T and 203 (Figure 2). PCR product from W. hellenica 4–7 was sequenced and the nucleotide sequences encoding the target gene was confirmed to be identical as reported (Figure 1). The results suggested that PCR amplification with the weissellicin L-specific primers could be useful to select potential weissellicin L-producing strain from multitudinous W. hellenica strains.Figure 2

Bottom Line: The obtained results indicated that the mature bacteriocin consists of 29 amino acid residues with a molecular weight of approximately 3205.64 Da.PCR presented a single product and was useful to detect weissellicin L structural gene.A rapid method to detect weissellicin L structural gene was also reported in this study.

View Article: PubMed Central - PubMed

Affiliation: Department of Biotechnology, Ming Chuan University, No. 5, De-Ming Rd., Gui-Shan Township, Taoyuan County, 333 Taiwan.

ABSTRACT

Background: Weissellicin L, a novel bacteriocin produced by Weissella hellenica 4-7, was previously characterized but its full amino acid sequence remain unknown. The draft genome sequencing analysis of Weissella hellenica 4-7 was performed and the open reading frame (ORF) encoding the weissellicin L was identified and clarified.

Findings: The obtained results indicated that the mature bacteriocin consists of 29 amino acid residues with a molecular weight of approximately 3205.64 Da. A conserved processing site of two glycine residues in positions -1 and -2 was observed in the leader peptides. The possibility that bacteriocin secretion depended on ATP-binding cassette (ABC) transporter was therefore suggested. Furthermore, primers were designed from 5' and 3' flanking sequences of the weissellicin L structural gene. PCR presented a single product and was useful to detect weissellicin L structural gene.

Conclusions: To our knowledge, this is the first report describing the full amino acid sequence of Weissellicin L. A rapid method to detect weissellicin L structural gene was also reported in this study.

No MeSH data available.