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Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis
Published on: October 31, 2014
Nucleotide sequence of the gene determining plasmid-mediated citrate utilization
Journal of Bacteriology
|December 1, 1985
Summary
Researchers cloned and sequenced the citrate utilization determinant from transposon Tn3411. They identified a potential gene encoding a hydrophobic polypeptide, but experimental results showed a smaller protein product.
Area of Science:
- Molecular Biology
- Genetics
- Microbiology
Background:
- Transposons are mobile genetic elements.
- Citrate utilization is an important metabolic pathway in bacteria.
Purpose of the Study:
- To clone and sequence the citrate utilization determinant from transposon Tn3411.
- To characterize the polypeptide products of this determinant.
Main Methods:
- DNA cloning and sequencing
- Minicell experiments
- Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)
Main Results:
- A 2,047-base-pair fragment containing the citrate determinant was sequenced.
- An open reading frame predicted a 431-amino-acid hydrophobic polypeptide.
- Minicell experiments revealed a single polypeptide with an apparent molecular weight of 35,000 Da.
Conclusions:
- The nucleotide sequence suggests a potential gene for citrate utilization.
- Experimental characterization indicates a discrepancy between the predicted and observed polypeptide size.
