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Molecular cloning, sequencing and expression of cDNA encoding human trehalase
R Ishihara1, S Taketani, M Sasai-Takedatsu
1Department of Pediatrics, Kansai Medical University, Osaka, Japan.
Gene
|January 14, 1998
Summary
Researchers isolated and expressed human trehalase cDNA, a brush-border membrane glycoprotein. This work facilitates understanding human trehalase gene structure and catalytic mechanisms.
Area of Science:
- Biochemistry
- Molecular Biology
- Genetics
Background:
- Trehalase is a glycoprotein found in brush-border membranes.
- Understanding human trehalase is crucial for various biological processes.
Purpose of the Study:
- To isolate and characterize the cDNA encoding human trehalase.
- To express the recombinant human trehalase and assess its activity.
- To provide a basis for further studies on human trehalase.
Main Methods:
- cDNA library screening (human kidney)
- Protein analysis (amino acid sequence, molecular weight, glycosylation sites)
- Northern blotting (mRNA expression)
- Recombinant expression in E. coli
Main Results:
- Isolated a complete cDNA clone for human trehalase (583 amino acids, 66,595 Da).
- Identified signal peptide, glycosylation sites, and membrane anchor region.
- Human trehalase sequence shows homology to other species' enzymes.
- mRNA predominantly expressed in kidney, liver, and small intestine.
- Recombinant enzyme exhibited high activity.
Conclusions:
- Successful isolation and expression of human trehalase cDNA.
- The findings facilitate research into human trehalase gene structure and function.
- This study provides a foundation for understanding the enzyme's catalytic mechanism.