Cloning And Analysis Of CDNA Encoding Odorant Binding Protein In The Antennae Of Adelphocoris Suturalis Jakovlev

Adelphocoris suturalis Jakovlev is one of the most destructive pests of cotton and cause significant yield losses every year. Adelphocoris suturalis Jakovlev can feed on a variety of plants, more than20families and80species. Between the opposite sex will communication by the insect sex pheromones, and further to mate and oviposition.In the process of insect sensitive olfactory system plays a very important role. Elucidating the mechanism of inseet olfactory perception can further facilitate the design and implementation of novel intervention strategies against these pests.In this paper,the function of odorant binding proteins in Adelphocoris suturalis Jakovlev was explored,and the main results are as follows:1. A575bp fragment, the complete cDNA sequence of OBP6including both the ORF459bp and3’end of the OBP6were respectively cloned using the homologous amplification and RACE method. The sequences encode153amino acids,termed AsuOBP6. The accession number is AHJ81241.1. The predicted MW and pI was17.5kDa and4.94,respectively.2. A748bp fragment, the complete cDNA sequence of OBP7including both the ORF603bp and3’end of the OBP7were respectively cloned using the homologous amplification and RACE method. The sequences encode201amino acids,termed AsuOBP7. The accession number is AHJ81239.1. The predicted MW and pI was22.4kDa and8.24,respectively.3. A645bp fragment, the complete cDNA sequence of OBP8including both the ORF450bp and3’end of the OBP8were respectively cloned using the homologous amplification and RACE method. The sequences encode150amino acids,termed AsuOBP8. The accession number is AHJ81242.1. The predicted MW and pI was16.9kDa and4.85,respectively.4. A682bp fragment, the complete cDNA sequence of OBP10including both the ORF483bp and3’end of the OBP10were respectively cloned using the homologous amplification and RACE method. The sequences encode161amino acids,termed AsuOBP10. The accession number is AHJ81240.1. The predicted MW and pI was18.3kDa and5.87,respectively.5. A594bp fragment, the complete cDNA sequence of OBP11including both the ORF450bp and3’end of the OBP11were respectively cloned using the homologous amplification and RACE method. The sequences encode150amino acids,termed AsuOBP11. The accession number is AHJ81243.1. The predicted MW and pI was16.8kDa and9.03,respectively.The first22amino acids was the signal peptide.6. A620bp fragment, the complete cDNA sequence of OBP12including both the ORF420bp and3’end of the OBP12were respectively cloned using the homologous amplification and RACE method. The sequences encode140amino acids,termed AsuOBP12. The accession number is AHJ81244.1. The predicted MW and pI was16.0kDa and5.27,respectively.7. We choose β-actin as the endogenous control to normalize the target gene expression and correct for sample-to-sample variation, choose AsuOBP6,AsuOBP8,AsuOBP11and AsuOBP12gene as the target gene, use the method of qPCR to study the relatively expression level of AsuOBP6,AsuOBP8,AsuOBP11and AsuOBP12gene in different development stages (1st,2nd,3rd,4th,5th) and different tissues (antenna, heads, thorax, abdomen, legs, wings). When the amplification efficiency between the target gene and the reference gene under the same, we use the comparative2-ΔΔCTmethod calculated the relative quantification of AsuOBP6,AsuOBP8.AsuOBP11and AsuOBP12directly. The result showed that AsuOBP6, AsuOBP8, AsuOBPll and AsuOBP12were mainly expressed at female antenna2nd of Adelphocoris suturalis Jakovlev, so we speculate that AsuOBP6, AsuOBP8, AsuOBPll and AsuOBP12play an important role in the female antenna of Adelphocoris suturalis Jakovlev. While AsuOBP6, AsuOBP8,AsuOBP11and AsuOBP12expressed in different development stages.