EXPRESSION AND PURIFICATION OF GLYOXYLATE/HYDROXY PYRUVATE REDUCTASE FROM BACILLUS SUBTILIS

Abstract

Background/Purpose: Gene expression refers to the process of translating genetic
information from Deoxyribonucleic Acid (DNA) to Ribonucleic Acid (mRNA) of a “special”
gene segment in a genome into a fully functional protein. Studying the function of a gene
is an important and meaningful job to better understand each gene segment’s function.
Glyoxylate/Hydroxypyruvate Reductase (GR/HPR) is a D-2-hydroxy-acid dehydrogenase
related to renal failure. The expression and purification of this gene play a very important
role in the research process, this is the first step to be able to conduct further studies on the
function and dynamics of the gene or protein that this gene encodes. Driving opportunities
for biotechnology uses in medicine development and medical treatment.
Methods: Described the experiments in the laboratory, gene transfer in bacteria, expression,
and purification of protein.
Results: We successfully cloned and expressed the gene encoding glyoxylate/hydroxypyruvate
dehydrogenase. The coding gene was inserted into the pET26b gene vector for the replicated
purpose in the cell and to collect in large quantities. After that, the transgenic vector was
inserted into the bacterial E. coli (DE3). Finally, from 2 liters of cell culture, we obtained 5
ml of recombinant protein solution concentration of 0.5mM (18.9 mg/ml).
Conclusion: We have successfully expressed the glyoxylate/hydroxypyruvate dehydrogenase
genes from Bacillus Subtillis (BsGOR). As well as the process of expression and purification
of this protein is also fully described in the study. Achieve the aim of research objectives.