Cloning of a cDNA encoding phosphofructokinase from Haemonchus contortus.
Mol Biochem Parasitol
; 48(1): 17-26, 1991 Sep.
Article
em En
| MEDLINE
| ID: mdl-1838137
ABSTRACT
Phosphofructokinase (PFK), the key regulatory enzyme in glycolysis, has been cloned from the pathogenic parasitic nematode Haemonchus contortus by functional complementation in Escherichia coli. An E. coli strain deleted for both PFK loci (strain DF1020) was transformed with plasmid DNA from a lambda ZAP II H. contortus cDNA library. Two out of 3 x 10(7) transformants were able to grow on minimal medium with mannitol as the sole carbon source. A plasmid, pPFK, containing a 2.7-kb insert, was isolated from one of these transformants and conferred on DF1020 the ability to grow on mannitol (the PFK phenotype). The complemented cells contain PFK enzyme activity, absent in the E. coli mutant, at levels considerably higher than in wild type E. coli. Sequence analysis of the 2.7-kb insert shows an open reading frame that predicts a 789-amino acid protein that has approximately 70% similarity to mammalian PFKs. The amino acid sequence around asp182, thought to be the catalytic site, is completely conserved from nematodes to mammals.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
DNA
/
Fosfofrutoquinase-1
/
Haemonchus
Limite:
Animals
Idioma:
En
Ano de publicação:
1991
Tipo de documento:
Article