Cloning and expression of isocitrate lyase from human round worm Strongyloides stercoralis.
A full length cDNA (1463 bp) encoding isocitrate lyase (EC 4.1.3.1) of Strongyloides stercoralis is described. The nucleotide sequence of this insert identified a cDNA coding for the isocitrate lyase. The conceptually translated amino acid sequence of the open reading frame for S. stercoralis isocitrate lyase encodes a 450 amino acid residue protein with an apparent molecular weight of 50 kDa and a predicted pl of 6.39. The sequence is 69% A/T, reflecting a characteristic A/T codon bias of S. stercoralis. The amino acid sequence of S. stercoralis isocitrate lyase is compared with bifunctional glyoxylate cycle protein of Caenorhabditis elegans and isocitrate lyases from Chlamydomonas reinhardtii and Myxococcus xanthus. The full length cDNA of S. stercoralis was expressed in pRSET vector and bacteriophage T7 promoter based expression system. S. stercoralis lyase recombinant protein, purified via immobilized metal affinity chromatography, showed a molecular mass of 50 kDa on polyacrylamide gels. The role of isocitrate lyase in the glyoxylate cycle and energy metabolism of S. stercoralis is also discussed.
Siddiqui
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Department of Internal Medicine, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, USA. siddiqui@etsu.edu
Stanley
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CS
Berk
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000201:enzymology; D016825:Chlamydomonas reinhardtii / Q000201:enzymology; D003001:Cloning, Molecular; D016384:Consensus Sequence; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007522:Isocitrate Lyase / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D017171:Strongyloides stercoralis / Q000201:enzymology*
D007522:Isocitrate Lyase