|Citation||Xu, Y., Intra, J., Zhang, C.X., Pasini, M.E. (2011). Recombinant expression of Drosophila melanogaster α-l-fucosidase in Trichoplusia ni cells. J. Insect Physiol. 57(9): 1205--1211. (Export to RIS)|
|Publication Type||Research paper|
|PubMed Abstract||A cDNA encoding an α-l-fucosidase from Drosophila melanogaster was obtained from the recombinant plasmid named pGEM-DmFuca and inserted into the pBacHTeGFPT vector to construct the recombinant donor plasmid which was transposed to the target AcBacmid in Escherichia coli (DH10) by Tn7 transposition function. The AcBacmid-GFP-DmFuca plasmid was used to transfect Tn-5B1-4 cells of the Cabbage looper Trichoplusia ni. SDS-PAGE analysis revealed a band of about 80kDa. Using a polyclonal antiserum raised against α-l-fucosidase protein from D. melanogaster Western blotting analysis confirmed that the fusion protein eGFP-DmFuca has been successfully expressed in a biologically active form in Tn-5B1-4 cells. The recombinant protein, containing the histidine-tag motif, was purified using an affinity chromatography column. In vitro binding assays the purified eGFP-DmFuca interacts with α-l-fucose residues present on the micropyle of the D. melanogaster eggshell, confirming that the α-l-fucosidase is a good candidate as receptor involved in gamete interactions in fruit fly.|
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|Language of Publication||English|
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|Also Published As|
|Abbreviation||J. Insect Physiol.|
|Title||Journal of Insect Physiology|
|Data from Reference|