AI Article Synopsis
- A plasmid shuttle vector was created using an E. coli plasmid, a bacteriophage D29 fragment, and a kanamycin resistance gene to efficiently introduce DNA into mycobacteria.
- The resulting plasmid, which is 7.63 kb in size, shows stability and exists in multiple copies within Mycobacterium smegmatis.
- The D29 fragment is believed to contain an origin of replication, with certain mutations impacting its ability to replicate inside M. smegmatis.
Article Abstract
A plasmid shuttle vector for Escherichia coli and mycobacteria was constructed from an E. coli plasmid containing the ColE1 origin, a 2.6-kb PstI fragment from bacteriophage D29 that grows in numerous mycobacterial species, and the kanamycin resistance gene either of Tn903 or of Tn5. The resultant plasmid is 7.63 kb and can be introduced via transformation into Mycobacterium smegmatis with high efficiency. In M. smegmatis the plasmid is stable and apparently present in multiple copies. Bioluminescence (luxA and luxB of Vibrio harveyi and fischeri) has been expressed in M. smegmatis from the aminoglycoside transferase promoter of Tn5. The D29 fragment should carry an origin of replication and some associated genes that act on it since various mutations destroy the ability of this fragment to replicate in M. smegmatis. The fragment was localized on the D29 genome map.
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| http://dx.doi.org/10.1016/0147-619x(92)90059-j | DOI Listing |
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