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...

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Bibliographic Details
Published in:Journal of medical entomology 2006-07, Vol.43 (4), p.707-712
Main Authors: Temeyer, Kevin B., Pruett, John H., Untalan, Pia M., Chen, Andrew C.
Format: Article
Language:English
Subjects:
Acari
acetylcholinesterase
Acetylcholinesterase - biosynthesis
Acetylcholinesterase - chemistry
Acetylcholinesterase - genetics
Acetylcholinesterase - metabolism
Animals
Arachnid Vectors - enzymology
Arachnid Vectors - genetics
Arachnid Vectors - physiology
Baculoviridae - genetics
baculovirus expression system
Benzenaminium, 4,4'-(3-oxo-1,5-pentanediyl)bis(N,N-dimethyl-N-2-propenyl-), Dibromide - pharmacology
Boophilus microplus
Cattle
Cholinesterase Inhibitors - pharmacology
Cloning, Molecular - methods
complementary DNA
DNA Primers - chemistry
DNA, Complementary - physiology
enzyme activity
enzyme inhibition
enzyme inhibitors
enzyme kinetics
gene expression
Gene Expression Regulation, Enzymologic - physiology
Insecticide Resistance - genetics
Insecticide Resistance - physiology
Ixodidae
Ixodidae - enzymology
Ixodidae - genetics
Ixodidae - physiology
kinetics
MOLECULAR BIOLOGY/GENOMICS
molecular cloning
nucleotide sequences
Paraoxon - pharmacology
Physostigmine - pharmacology
recombinant proteins
substrate specificity
tick
ticks
Transfection
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Description
Summary: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.
ISSN:0022-2585
1938-2928
DOI:10.1603/0022-2585%282006%2943%5B707%3ABEOBAC%5D2.0.CO%3B2