What question did this study set out to answer?

To investigate the feasibility, frequency, and genomic characteristics of transposon Tn5 insertion mutagenesis in plant-pathogenic Erwinia carotovora subspecies using the suicide plasmid pJB4JI.

March 1, 1984Journal of BacteriologyOpen Access

Transposon Tn5 mutagenesis in Erwinia carotovora subsp. carotovora and E. carotovora subsp. atroseptica

View Full Paper

Ask AI

Bookmark

Authors

RZRalf ZinkCognizant (United States)RKR. J. KembleKansas State University ACArun ChatterjeeThapar Institute of Engineering & Technology

Discussion

Member takes

Implication

Experimental study demonstrates high-frequency Tn5 insertion mutagenesis in Erwinia carotovora subspecies, indicating its efficacy for isolating diverse metabolic mutants.

Key Points

To investigate the feasibility, frequency, and genomic characteristics of transposon Tn5 insertion mutagenesis in plant-pathogenic Erwinia carotovora subspecies using the suicide plasmid pJB4JI.
Conjugative mating between Escherichia coli 2492 harboring plasmid pJB4JI and recipient strains Erwinia carotovora subsp. carotovora Ecc71 and E. carotovora subsp. atroseptica Eca12.
Phenotypic screening of kanamycin-resistant, gentamicin-sensitive (Kmr Gms) transconjugants to identify auxotrophic mutants and carbohydrate utilization defects.
Southern blot hybridization with 32P-labeled pRZ102 probe DNA to verify Tn5 insertion and confirm the absence of bacteriophage Mu plasmid sequences.
Transconjugants with a Kmr Gms phenotype emerged at high frequencies, with 2% to 3% exhibiting auxotrophy or defective catabolism of specific carbohydrates.
Southern blot hybridization confirmed that phenotype alterations resulted predominantly from single, scattered insertions of Tn5 into the host chromosome without accompanying Mu DNA.
Spontaneous prototrophic revertants were successfully recovered for most phenotypic markers, with the exception of ilv, tyr, and suc.

Cite This Study

Zink et al. (1984) studied this question.

synapsesocial.com/papers/6a8a9ae04c30e1095aecc02d https://doi.org/10.1128/jb.157.3.809-814.1984

View Full Paper

Ask AI

Bookmark

March 1, 1984Journal of BacteriologyOpen Access

Transposon Tn5 mutagenesis in Erwinia carotovora subsp. carotovora and E. carotovora subsp. atroseptica

View Full Paper

Ask AI

Bookmark

Authors

RZRalf ZinkCognizant (United States)RKR. J. KembleKansas State University ACArun ChatterjeeThapar Institute of Engineering & Technology

Discussion

Member takes

Implication

Experimental study demonstrates high-frequency Tn5 insertion mutagenesis in Erwinia carotovora subspecies, indicating its efficacy for isolating diverse metabolic mutants.

Key Points

To investigate the feasibility, frequency, and genomic characteristics of transposon Tn5 insertion mutagenesis in plant-pathogenic Erwinia carotovora subspecies using the suicide plasmid pJB4JI.
Conjugative mating between Escherichia coli 2492 harboring plasmid pJB4JI and recipient strains Erwinia carotovora subsp. carotovora Ecc71 and E. carotovora subsp. atroseptica Eca12.
Phenotypic screening of kanamycin-resistant, gentamicin-sensitive (Kmr Gms) transconjugants to identify auxotrophic mutants and carbohydrate utilization defects.
Southern blot hybridization with 32P-labeled pRZ102 probe DNA to verify Tn5 insertion and confirm the absence of bacteriophage Mu plasmid sequences.
Transconjugants with a Kmr Gms phenotype emerged at high frequencies, with 2% to 3% exhibiting auxotrophy or defective catabolism of specific carbohydrates.
Southern blot hybridization confirmed that phenotype alterations resulted predominantly from single, scattered insertions of Tn5 into the host chromosome without accompanying Mu DNA.
Spontaneous prototrophic revertants were successfully recovered for most phenotypic markers, with the exception of ilv, tyr, and suc.

Cite This Study

Zink et al. (1984) studied this question.

synapsesocial.com/papers/6a8a9ae04c30e1095aecc02d https://doi.org/10.1128/jb.157.3.809-814.1984

Also Consider

Synapse has enriched 2 closely related papers on similar clinical questions. Consider them for comparative context:

View Full Paper

Ask AI

Bookmark