中华蜜蜂化学感受蛋白基因Acer-CSP1克隆与表达特征分析

Molecular cloning,characterization,and expression pattern of chemosensory protein 1 gene(Acer-CSP1)in the Chinese honeybee,Apis cerana cerana(Hymenoptera:Apidae)

化学感受蛋白（chemosensory proteins,CSPs）是昆虫化学感受系统中重要的组成部分之一。本研究克隆了中华蜜蜂Apis cerana cerana化学感受蛋白基因Acer-CSP1,其核苷酸全长351bp（GenBank登录号为FJ157352）,编码116个氨基酸残基,预测蛋白分子量为13.85kD,等电点为4.89,且含有4个保守的半胱氨酸残基,均符合昆虫CSPs的一般特征,且与意蜂CSP1基因具有99.1%的相似性,与其他昆虫也有45.3%~68.0%的相似性。利用2-ΔΔCt法及绝对定量法的real-time PCR技术对Acer-CSP1在中蜂不同器官表达特征进行了研究,得出的一致结论为Acer-CSP1显著水平地高丰度表达于中华蜜蜂触角,其次大量表达于头部。由于触角为中华蜜蜂最主要的嗅觉器官,而头部则具有发达的感觉神经系统和味觉系统,这也提示Acer-CSP1极有可能参与中华蜜蜂的嗅觉以及其他化学感受功能。

Chemosensory proteins（CSPs）are a class of the most important proteins involved in chemical communications and perception in insects.In this study,one CSP gene from the Chinese honeybee,Apis cerana cerana,was cloned and characterized,which was named Acer-CSP1（GenBank accession no.：FJ157352）.The Acer-CSP1 cDNA was 351 bp in length,encoding 116 amino acid residues with the predicated molecular mass of approximately 13.85 kD and pI of 4.89,of which four conservative cysteines are included.With the general characteristics of the CSP gene family in insects,Acer-CSP1 showed high similarity（99.1%）with the CSP1 gene of A.mellifera and moderate similarity（45.3%-68.0%）with the CSP genes of other ten insects analyzed with the deduced amino acid sequences.The expression of the Acer-CSP1 in different sensory organs of A.cerana cerana was compared using both relative（2-ΔΔCt）and absolute quantitative analyses of real-time PCR.The results indicated that the Acer-CSP1 was primarily expressed in the antennae,followed by the head,but least expressed in other organs（wings,legs,thorax,and abdomen）.Since the antennae are the major olfactory organs and the head is the major sensory and gustatory organ of the honeybee,the results suggest that the Acer-CSP1 gene is most likely involved in the olfaction,gestation and other chemosensory functions of the insect.