Abstract: 【Objective】 The objective of this study is to clone the full-length cDNA sequence of the cAMP response element binding protein gene of Apis cerana cerana (AcCREB), predict the gene and its physicochemical properties of the corresponding encoded protein, and then clarify its expression in the brain tissues, which will provide a research basis of the physiological function of AcCREB in the learning and memory. 【Method】Total RNA was extracted from the dissected brain tissues of A. cerana cerana (A.c.c.) as the main experimental materials. On this basis, the AcCREB was cloned using RT-PCR technique. The forward and reverse sequences were then spliced by Bioedit to obtain the full-length cDNA using the Seqman of DNAstar software, and its amino acid sequences were translated using Bioedit software according to the six-box translation. The sequences was analyzed using BLASTn and BLASTp. Multiple sequence alignment and homology were analyzed using ClustalX. A phylogenetic tree was constructed using neighbor-joining method of software MEGA. The conserved structural domains were deduced by analyzing the SMART database. In addition, the paraffin sections of brain were made based on the dissected brain tissues, and the distribution of its expression in the brain was investigated by in situ hybridization histochemistry. 【Result】The full length of AcCREB cDNA is 890 bp, which encodes 240 amino acids. The sequence was submitted to GenBank (Accession number KC814690). The molecular weight and an isoelectric point of AcCREB protein were 25.691 kD and 5.82, respectively. Structural domain analysis indicated that the encoded protein contained two highly conserved structures of AA 89-131 PKID and AA 176-237 BRLA. Homology comparison illustrated that AcCREB has a similarity with CREB of Apis mellifera (AmCREB), Bombus impatiens (BiCREB), Megachile rotundata (MrCREB), Bombus terrestris (BtCREB), Harpegnathos saltator (HsCREB), Bombyx mori (BmCREB), Aedes aegypt (AaCREB),Anopheles gambiae (AgCREB), Homo sapiens (HsCREB)and Mus musculus (MmCREB), with the similarity of 98.76%, 98.35%, 97.11%, 94.09%, 72.22%, 58.97%, 44.69%, 41.69%, 36.89%, and 36.89%, respectively. Meanwhile, the phylogenetic tree analysis showed that AcCREB was first clustered with AmCREB, BiCREB, BtCREB. Moreover, in situ hybridization showed that AcCREB staining was mainly localized to the Kenyon cells of the mushroom bodies, cells around the antennal lobes, cells between medulla and lobula, and display asymmetrical expression between the right and left brain of worker bees in A.c.c..【Conclusion】The amino sequences of AcCREB own high homology with AmCREB, BiCREB, MrCREB, BtCREB, and the distribution of the gene in the brains of honeybee suggested that AcCREB may be involved in the process of learning and memory of the A. c. c. which provides useful information for future studies of CREB in the leaning-memory of honeybees.

Key words: Apis cerana cerana, cAMP response element binding protein gene, molecular cloning, expression analysis