Related Experiment Videos
Studies on Tn951 (lac+) expression in Agrobacterium
Molecular & General Genetics : MGG
|January 1, 1984
Summary
Agrobacterium tumefaciens and A. rubi strains do not produce beta-galactosidase. Introducing the lac transposon Tn951 enabled these bacteria to produce this enzyme, facilitating alien gene expression studies.
Area of Science:
- Microbiology
- Molecular Biology
- Bacteriology
Background:
- Agrobacterium tumefaciens and A. rubi strains utilize lactose but lack detectable beta-galactosidase activity.
- Investigating alien gene expression in Agrobacterium is crucial for genetic engineering applications.
Purpose of the Study:
- To investigate the expression of the lac transposon Tn951 in Agrobacterium species.
- To establish a system for studying alien gene expression in Agrobacterium.
Main Methods:
- Introduction of the lac transposon Tn951 into Agrobacterium using the broad-host range plasmid RP1.
- Assessing beta-galactosidase production in transconjugants.
- Testing the ability of Tn951 to restore lactose utilization (Lac+ phenotype) in mutant strains.
Main Results:
- Transconjugants carrying Tn951 produced significant quantities of beta-galactosidase, inducible by isopropyl-beta-D-thiogalactopyranoside.
- Tn951 successfully restored the Lac+ phenotype to an Agrobacterium tumefaciens mutant.
- Cellobiose, a known inducer of a related enzyme, did not affect beta-galactosidase activity.
Conclusions:
- The lac transposon Tn951 can be effectively expressed in Agrobacterium species.
- Tn951 provides a functional system for studying gene expression in Agrobacterium.
- This system holds potential for investigating alien gene expression in these bacteria.