关键词: 中华蜜蜂, 工蜂触角, 气味结合蛋白, 原核表达

Abstract: 【OBJECTIVE】Cloning and expression in prokaryotic system of a new cDNA, named Acer-ASP2, encoding odorant binding protein ASP2(antenna special protein 2) open reading frame(ORF).【METHOD】Based on the antenna materials of 13 ages in days from Apis cerana cerana, reverse transcription-polymerase chain reaction(RT-PCR) and pET-30a(+)/BL21(DE3) prokaryotic expression system, Acer-ASP2 was cloned and expressed. 【RESULTS】The full length of Acer-ASP2(GenBank loucs: DQ449667) was 429bp, encoding 142 amino acids and the predicted MW and pI were 15.7 kD and 4.36, respectively. Sequencing and analysis indicated that Acer-ASP2 was characterized by six conservative Cys, which shared typical feature of OBP from other insects. Further analysis showed Acer-ASP2 likely belongs to the family of GOBP. The molecular weight of recombinant protein of pET/Acer-ASP2 was about 21kD, induced by IPTG to highly express a protein of as an inclusion body and accumulated up to 59.7% of bacterial total protein.【CONCLUSION】We have cloned and expressed a new cDNA of Apis cerana cerana, for the further research to the molecular structure and function.

Key words: Apis cerana cerana, Working bee´, s antenna, Odorant Binding Protein, Prokaryotic express