Cloning, expression and characterisation of a recombinant triosephosphate isomerase from Taenia solium.
We isolated and characterised the cDNA that encodes the glycolytic enzyme, triosephosphate isomerase from Taenia solium. A 450 bp DNA fragment was obtained by the polymerase chain reaction using a cDNA from larval stage as template and degenerate oligonucleotides designed from conserved polypeptide sequences from TPIs of several organisms. The fragment was used to screen a T. solium larval stage cDNA library. The isolated cDNA, encoding a protein of 250 amino acids shares 44.8-59.6% positional identity with other known TPIs, in which the catalytic enzyme residues were conserved. The complete coding sequence of the T. solium TPI cDNA was cloned into the expression vector pRSET and expressed as a fusion protein with an N-terminal tail of six histidine residues. The catalytic activity of the purified protein was similar to other TPI enzymes. Northern and Southern blot analysis suggest that in T. solium, single gene exists for triosephosphate isomerase and that the gene is expressed in all stages of the parasite.
Jiménez
L
L
Departamento de Microbiología y Parasitología,Facultad de Medicina, Universidad Nacional Autónoma de México, Edificio A, 2 degrees Piso, CiudadUniversitaria, Mexico D.F. 04510, México.
Vibanco-Pérez
N
N
Navarro
L
L
Landa
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D003551:Cysticercosis / Q000469:parasitology*; D017931:DNA Primers / Q000737:chemistry; D016054:DNA, Protozoan / Q000737:chemistry; D004926:Escherichia coli / Q000737:chemistry; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D016053:RNA, Protozoan / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013552:Swine; D013621:Taenia / Q000201:enzymology / Q000235:genetics*; D014305:Triose-Phosphate Isomerase / Q000737:chemistry / Q000235:genetics*
D017931:DNA Primers; D016054:DNA, Protozoan; D016053:RNA, Protozoan; D011994:Recombinant Proteins; D014305:Triose-Phosphate Isomerase