Cloning of the BamHI methyl transferase gene from Bacillus amyloliquefaciens |
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| Affiliation: | 1. Research Institute for Viticulture and Oenology, University of Pécs, Pázmány P. u. 4, Pécs H-7634, Hungary;2. Department of Plant Biology, University of Pécs, Ifjúság u. 6, Pécs H-7624, Hungary;3. Institute of Organic and Medicinal Chemistry, University of Pécs, Szigeti st. 12, H-7624 Pécs, Hungary;4. Electron Microscopy Facility, Istituto Italiano di Tecnologia, via Morego 30, Genova 16163, Italy;5. Department of Nanochemistry, Istituto Italiano di Tecnologia, via Morego 30, Genova 16163, Italy;1. Biochemical Adaptation Laboratory, Department of Zoology, North-Eastern Hill University, Shillong 793 022, India;2. Department of Biotechnology, Assam Don Bosco University, Assam 782402, India |
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| Abstract: | We wish to report the initial characterization of a recombinant clone containing the BamHI methylase gene. Genomic chromosomal DNA purified from Bacillus amyloliquefaciens was partially cleaved with HindIII, fractionated by size, and cloned into pSP64. Plasmid DNA from this library was challenged with BamHI endonuclease and transformed into Escherichia coli HB101. A recombinant plasmid pBamM6.5 and a subclone pBamM2.5 were shown to contain the BamHI methylase gene based on three independent observations. Both plasmids were found to be resistant to BamHI endonuclease cleavage, and chromosomal DNA isolated from E. coli HB101 cells harboring either of the plasmids pBamM6.5 or pBamM2.5 was resistant to cleavage by BamHI endonuclease. In addition, DNA isolated from lambda phage passaged through E. coli HB101 containing either plasmid was also resistant to BamHI cleavage. Expression of the BamHI methylase gene is dependent on orientation in pSP64. In these clones preliminary evidence indicates that methylase gene expression may be under the direction of the plasmid encoded LacZ promoter. |
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