A PCR strategy for the isolation of glutathione S-transferases (GSTs) from nematodes.
Anchor based PCR technology has been used to isolate a GST sequence from the gastro-intestinal nematode Heligmosomoides polygyrus. A 800 base pair product was amplified from first-strand cDNA using primers based on the N-terminal sequence of purified H. polygyrus GST (upstream primer) and a non-specific polyadenylate tail with an anchor sequence (downstream primer). The product was cloned into pUC18 and sequenced. A reading frame of 648 bases in the sequence encoded a protein which has 30% homology with the alpha family of mammalian glutathione S-transferases.
Brophy
P M
PM
Department of Pharmacy, School of Applied Sciences, De Montfort University, Leicester, U.K.
Brown
A
A
Pritchard
D I
DI
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D005982:Glutathione Transferase / Q000235:genetics*; D008969:Molecular Sequence Data; D009351:Nematospiroides dubius / Q000201:enzymology / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D005982:Glutathione Transferase