Cloning, sequencing and functional expression of a DNA encoding pig cytosolic malate dehydrogenase: purification and characterization of the recombinant enzyme.
Gene
; 172(2): 303-8, 1996 Jun 26.
Article
in En
| MEDLINE
| ID: mdl-8682322
ABSTRACT
Using the polymerase chain reaction, DNA encoding cytosolic malate dehydrogenase (cMDH) has been cloned from a pig heart cDNA library. Large amounts of the enzyme (30 mg per litre of original culture) have been produced in Escherichia coli using an inducible expression vector (pKK223-3) in which the 5'-non-coding region of the gene was replaced with the tac promoter. The complete nucleotide sequence of the DNA is reported for the first time. The recombinant cMDH purified was shown to be identical to the native enzyme according to chromatographic behaviour, isoelectric point, N-terminal amino acid sequence, and physiochemical and catalytic properties.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Malate Dehydrogenase
Limits:
Animals
Language:
En
Journal:
Gene
Year:
1996
Document type:
Article
Affiliation country:
España