Dorathy Anak Jampi (2018) Cloning and expression of putative antimicrobial coding genes from plasmid pMWHK1 of pedobacter cryoconitis strain BG5. Masters thesis, Universiti Malaysia Sabah.
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Abstract
Pedobacter cryoconitisstrain BGS harbours a plasmid, pMWHKl. The circular plasmid consists of 6206 bp nucleotides and eight open reading frames (ORFs) which are deduced as genes. P. cryoconitis BGS produces antimicrobial compounds that are able to inhibit the growth of food pathogens. However, to date it is not known whether the antimicrobial compounds are conferred by genes in the plasmid or chromosome. There is possibility that the antimicrobial compounds are encoded by genes in plasmid, pMWHKl. Four out of the eight open reading frames were selected in this study which are orf5, orf6, orf7and orfB. OrfSis hypothesized to encode for an antitoxin gene whereas orf6, orf7 and orfB are hypothesized to encode for the antimicrobial gene. The study was conducted to determine if antimicrobial properties of P. cryoconitis BGS are conferred by orf6, orf7 and orfB of plasmid pMWHKl. Besides, the study was set-up to determine if the antitoxin gene is encoded by orfS. Another objective of the study is to determine the toxin and antitoxin effect of the genes of plasmid pMWHKl. The putative genes were analysed by expressing them in £ coli. Following expression, the antimicrobial activities and proteins were assayed. Orf5, orf6, orf7 and orfB were successfully cloned and expressed. Among the ORFs tested, the protein encoded by orf6was able to inhibit the growth of Bacillus cereusK3 and reduce the growth rate of£ co/icarrying that gene. However as tested with expression vectors in £ coli host, orf5 was unable to show effect against orf6 and no toxin-antitoxin effect of the genes were observed. Hence, further study is required as the study may discover new antimicrobial compounds. Findings in this study provided basic foundation or understanding on plasmid pMWHKl.
| Item Type: | Thesis (Masters) |
|---|---|
| Keyword: | Pedobacter cryoconitis, Plasmid pMWHK1, Antimicrobial gene, Open reading frame |
| Subjects: | Q Science > QH Natural history > QH301-705.5 Biology (General) > QH426-470 Genetics |
| Department: | INSTITUTE > Biotechnology Research Institute (BRI) |
| Depositing User: | DG MASNIAH AHMAD - |
| Date Deposited: | 11 Jun 2025 11:18 |
| Last Modified: | 11 Jun 2025 11:18 |
| URI: | https://eprints.ums.edu.my/id/eprint/44026 |
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