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Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 27, No. 1, 2006, pp. 75-80
Bioline Code: zr06012
Full paper language: Chinese
Document type: Research Article
Document available free of charge

Zoological Research, Vol. 27, No. 1, 2006, pp. 75-80

 en Expression of the Aphis gossypii check for this species in other resources AChE1 Gene Using the Baculovirus Expression System
DONG Shuang-lin, HAN Zhao-jun, Martin S WILLIAMSON

Abstract

Using the Bac-to-Bac baculovirus expression system, the acetylcholinesterase 1 gene (ace 1) of Aphis gossypii check for this species in other resources was successfully expressed in Sf9 cells. The enzyme was secreted into the cell culture medium and tested. The affinity (Km) of the expressed AChE1 for the substrate analog ATChI was 144.5±28.6 μM. The sensitivity index (indicated by Ki value) of expressed AChE1 to 2 carbamate insecticides (pirimicarb and triazamate) and 2 organophosphate insecticides (omethoate and demeton-S-methyl) were 513, 1 883, 5 and 23 (mM·min)-1, respectively, which is in agreement with previously reported values obtained with the natural enzyme. The expressed enzyme showed highest activity when harvested at 2-3 d after infection of the insect cells. The expressed AChE1 was very stable without obvious loss in enzyme activity after 90 d when stored at -20 °C, although 70% of the enzyme activity was lost after 7 d when stored at room temperature.

Keywords
Bac-to-Bac baculovirus expression system; Aphis gossypii; Acetylcholenesterase; Sf9 cell strain

 
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