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Cloning, E. coli Expression, and Characterization of Heart Lactate Dehydrogenase B From River Buffalo (Bubalus bubalis).
MedLine Citation:
PMID:  24299182     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Lactate dehydrogenase is an enzyme of glycolytic pathway which catalyzes the interconversion of pyruvate and lactate. The present study describes cDNA cloning, E. coli expression and characterization of lactate dehydrogenase B (LDH-B) from the heart ventricles of river buffalo (Bubalus bubalis). Total RNA was isolated from the heart tissue, a 1005bp cDNA encoding complete polypeptide chain of 334 amino acids was generated by reverse transcriptase reaction and analyzed for nucleotide sequence. The consensus sequence obtained from both strands has shown 84% to 98% homology with that of different mammalian species. The attributed gene was cloned, expressed in BL21 (DE3) RIPL Codon Plus strain of E. coli using pET21a (+) plasmid. The purified recombinant enzyme displayed a KM value of 50 µM for pyruvate, an optimum activity at 35°C and pH 7.0. The enzyme was found as a homotetramer of 140 kDa on FPLC based gel-filtration column. Molecular weight of a subunit of enzyme as determined by mass spectrometric analysis was 36530.21 Da. The present study describes the first ever report about the cDNA sequence and characteristics of recombinant LDH-B from River buffalo.
Authors:
Muhammad Shahid Nadeem; Jenny Moran; Bibi Nazia Murtaza; Khushi Muhammad; Habib Ahmad
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Animal biotechnology     Volume:  25     ISSN:  1532-2378     ISO Abbreviation:  Anim. Biotechnol.     Publication Date:  2014 Jan 
Date Detail:
Created Date:  2013-12-04     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9011409     Medline TA:  Anim Biotechnol     Country:  England    
Other Details:
Languages:  eng     Pagination:  23-34     Citation Subset:  IM    
Affiliation:
a Department of Genetics , Hazara University, Garden Campus , Mansehra , Pakistan.
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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