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Cloning and sequencing of a jack bean urease-encoding cDNA
P W Riddles1, V Whan, R L Blakeley
1CSIRO, Division of Tropical Animal Production, Long Pocket Laboratories, Indooroopilly, Queensland, Australia.
Gene
|December 15, 1991
Summary
Researchers cloned the complementary DNA (cDNA) for jack bean urease, determining its complete amino acid sequence. This marks the first report of cloned urease cDNA from a higher plant.
Area of Science:
- Biochemistry
- Molecular Biology
- Plant Science
Background:
- Urease is a crucial enzyme in plants.
- The genetic basis of jack bean urease was previously uncharacterized.
Purpose of the Study:
- To clone and sequence the complementary DNA (cDNA) encoding mature jack bean urease.
- To determine the complete amino acid sequence of jack bean urease.
Main Methods:
- Construction of a complementary DNA (cDNA) library from developing jack bean mRNA.
- Use of reverse transcriptase to obtain cDNA fragments.
- Complete sequencing of two contiguous DNA fragments.
- Conceptual translation of nucleotide sequence to amino acid sequence.
Main Results:
- The entire amino acid sequence of mature jack bean urease was determined.
- The cDNA sequence yielded an 840-amino acid sequence.
- The deduced amino acid sequence was identical to the directly determined sequence, with one conservative substitution.
Conclusions:
- This study reports the first cloning and sequencing of urease cDNA from any higher plant.
- The findings provide a molecular basis for understanding jack bean urease structure and function.