J Korean Soc Microbiol.
1997 Jun;32(3):293-300.
Construction of a Mycobacterium - Escherichia coli Shuttle Vector and Use in the Expression of Foreign Genes in Mycobacteria
Abstract
- The ability to introduce recombinant DNA molecules back into mycobacteria would greatly increase the potential of molecular genetic approaches for the study of mycobacteria as well as for the use in clinical purposes. We have initiated the construction of vectors that facilitates the introduction of recombinant DNA into mycobacteria. The vector was designed to contain replicons for multiplication in mycobacteria and Escherichia coli, a promoter for gene expression, a drug resistant gene for selecting transformants, and a few restriction enzyme sites for convenient cloning. Constructed Mycobacterium-E. coli shuttle vector named p YMC (hsp60) was shown to transform M. smegmatis at high efficiency and maintain plasmid at stable level. The ability of the vector to express cloned foreign gene was also monitored by measuring the expressed level of luciferase gene which was used as a reporter. High level of luciferase activity in M. smegmatis with pYMC (hsp60:luc) was detected confirming successful construction of Mycobacterium-E. coli shuttle vector.
- Full Text Links
-
- Actions
-
Cited
- CITED
-
- Close
- Share
-
- Similar articles
-
- Development of a Mycobacterial Gene Knock-out System using Sequence-specific Recombinase FLP/FRT and its Application to the Construction of a Rhamnose Biosynthetic Gene rmlD Deletion Mutant
- Expression of the 38 kDa Protein of Mycobacterium tuberculosis in M . bovis BCG and Use in the Serodiagnosis of Tuberculosis
- Factors affecting transformation efficiency of BCG with a Mycobacterium-Escherichia coli shuttle vector pYUB18 by electroporation
- Expression of human CTL?4 extracellular domain in escherichia coli
- Up-regulation of prothymosin alpha in THP-1 cells infected with Mycobacterium tuberculosis
