关键词: 中华蜜蜂 , 感觉神经元膜蛋白 , 基因克隆 ,  组织表达模式 , 原核表达

Abstract:
To explore the signal transduction pathway of important factors in olfactory formation in Apis cerana cerana, the cDNA sequence encoding a sensory neuron membrane protein (SNMP) (GenBank accession no. KC012595), named as AccSNMP1, was cloned by RT-PCR from the Chinese honey bee, Apis cerana cerana. Sequence analysis results showed that the open reading frame (ORF) is 1 563 bp in length, encoding 520 amino acids with the predicted molecular weight of 58.02 kD and the theoretical isoelectric point of 5.83. Multiple sequence alignment indicated that AccSNMP1 from A. cerana cerana shares different identities with those from other nine insects at the amino acid level. The AccSNMP1 gene from A. cerana cerana has high amino acid sequence identity with that of Apis mellifera (99.2%) and Bombus impatiens (90.9%), while has the lowest amino acid sequence identity with that of Tribolium castaneum (22.7%). The phylogenetic analysis indicated that A. cerana cerana species has a close relationship with A. mellifera and Bombus impatiens. Tissue expression profiling of AccSNMP1 quantified by real time RT-PCR demonstrated that it was highly expressed in antennae and legs of A. cerana cerana, showing a significant difference with that in thorax, abdomen, proboscis and head without antennae and proboscis (P<0.05). A recombined plasmid pEASY-E1-AccSNMP1, containing the coding sequence of AccSNMP1, was constructed using pEASY-E1 as the fused expression vector, and AccSNMP1 was expressed successfully after induced with IPTG in BL21 (DE3) strain of Escherichia coli. The results provide the basis for further studying the functions of sensory neuron membrane protein gene in A. cerana cerana to better understand its action mechanisms.

Key words: Apis cerana cerana , sensory neuron membrane protein , gene cloning , tissue expression profile , prokaryotic expression