Baculovirus expression of BmAChE3, a cDNA encoding an acetylcholinesterase of Boophilus microplus (Acari: Ixodidae).
The complete cDNA sequence encoding a Boophilus microplus (Canestrini) (Acari: Ixodidae) acetylcholinesterase (AChE3) was expressed in the baculovirus system. The recombinant AChE3 protein (rBmAChE3) was secreted as a soluble form into the cell culture medium and was identified as a functional AChE by substrate specificity and by inhibition with the AChE-specific inhibitors eserine sulfate and BW284c51. Inhibition kinetics of rBmAChE3, in the presence of the organophosphate paraoxon, revealed sensitivity comparable with that of adult, organophosphate-susceptible neural AChE. To our knowledge, this is the first report of the cloning and successful expression of a functional ixodid AChE.
Temeyer
Kevin B
KB
Knipling-Bushland U.S. Livestock Insects Research Laboratory, USDA-ARS, 2700 Fredericksburg Road, Kerrville, TX 78028, USA. kevin.temeyer@ars.usda.gov
Pruett
John H
JH
Untalan
Pia M
PM
Chen
Andrew C
AC
D000110:Acetylcholinesterase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001096:Arachnid Vectors / Q000201:enzymology / Q000235:genetics / Q000502:physiology; D016367:Baculoviridae / Q000235:genetics; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide / Q000494:pharmacology; D002417:Cattle; D002800:Cholinesterase Inhibitors / Q000494:pharmacology; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D007305:Insecticide Resistance / Q000235:genetics / Q000502:physiology*; D026863:Ixodidae / Q000201:enzymology* / Q000235:genetics / Q000502:physiology*; D010261:Paraoxon / Q000494:pharmacology; D010830:Physostigmine / Q000494:pharmacology; D014162:Transfection
D002800:Cholinesterase Inhibitors; D017931:DNA Primers; D018076:DNA, Complementary; D002093:Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide; D010830:Physostigmine; D000110:Acetylcholinesterase; D010261:Paraoxon