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| Cloning and Expression of Gene Encoding Gqα-protein in the England Grain Aphid (Sitobion avenae ) |
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Abstract Based on conserved homologous amino acid sequences of Gq protein α subunit in arthropods, a pair of degenerate primers were designed to amplify the gene from the England grain aphid(Sitobion avenae) by RT-PCR and (3′/5′)-RACE techiques. A Gqα was obtained from the alate adult aphids. The open reading-frame was 1 062 bp, encoding 352 amino acid residues with calculated molecular weight of 40.8 kD. The cDNA sequence was deposited in GenBank with the accession number EF638906. The deduced amino acid sequence of Gqα shared high identity (≥82.17%) with reported Gqα from other insects and even vertebrates, and with the typical characteristics of Gqα protein. In order to explore the function of Gqα gene, a eukaryotic expressional system(Baculovirus expression vector system, BEVS) was constructed by TOPO and Gateway technique. After the recombinant reaction between pUC-Gqα and Gateway-adapted Baculovirus DNA from Autographa californica nuclear polyhedrosis virus (AcMNPV), the constructed recombinant viruses containing V5-His6Gqα were transfected singly into an insect (Trichoplusia ni) cell line of Tn-5B1-4. After collecting the infected cell, an expected protein (about 42 kD) was detected by SDS-PAGE and Western blot. The results showed that the system of recombinant Baculovirus and Tn could express Gqα protein successfully.
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Received: 14 March 2008
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