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Published on: February 5, 2021
Functional analysis of pAL5000, a plasmid from Mycobacterium fortuitum: construction of a "mini"
M G Ranes1, J Rauzier, M Lagranderie
1Unité de Génie Microbiologique, Institut Pasteur, Paris, France.
Abstract:
Functional domains of pAL5000 were determined by gene disruption and deletion analysis. Of the five plasmid open reading frames (ORFs), ORF1 to ORF5, and a putative origin of replication previously identified (J. Rauzier, J. Moniz-Pereira, and B. Gicquel-Sanzey, Gene 71:315-321), two of the ORFs (ORF3 and ORF4) were deemed dispensable for plasmid replication. A "mini" mycobacterium-Escherichia coli shuttle plasmid applicable for general recombinant DNA studies in mycobacteria was constructed by using the gene for Kanr (Tn903) as a selective marker. Heterologous expression of the gene for Kanr was confirmed by Western blotting (immunoblotting) analysis.
