Cloning and Expression of Pseudomonas aeruginosa AlkB Gene in E. coli
Pre identified hydrocarbons degrading bacteria were used in this study, specific primer was conducted to amplification of AlkB gene, approximately 1206bp band size of this gene for Pseudomonas aeruginosa was detected and proofed by sequence and alignment analysis with NCBI database. The AlkB gene...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Journal of Pure and Applied Microbiology
2020-03-01
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| Series: | Journal of Pure and Applied Microbiology |
| Subjects: | |
| Online Access: | https://microbiologyjournal.org/cloning-and-expression-of-pseudomonas-aeruginosa-alkb-gene-in-e-coli/ |
| Summary: | Pre identified hydrocarbons degrading bacteria were used in this study, specific primer was conducted
to amplification of AlkB gene, approximately 1206bp band size of this gene for Pseudomonas aeruginosa
was detected and proofed by sequence and alignment analysis with NCBI database. The AlkB gene was
inserted in PET-21a(+) plasmid vector as expression vector, then transformed in BL21(DE3) competent
E. coli and confirmed by colony PCR technique using the T7 promoter and T7 terminator primers. The
expression of the inserted gene was checked by determined the concentration of AlkB protein for
multiple periods by Bradford assay method and the SDS-polyacrylamide gel electrophoresis method
was revealed band of ~46 KD molecular weight of the concerned protein. The gene amplification and
cloning strategy was lay out before the practical part of the study by SnapGene software, this study was
conducted to introduce cloned bacteria which facilitate the first step (key step) of alkane’s biodegradation
and propose an appropriate strategy to construct genetically engineered microorganisms with multiple
recombinant plasmid for enhance the degradation of the aliphatic fraction of hydrocarbon |
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| ISSN: | 0973-7510 2581-690X |