| 蜜蜂表皮蛋白apidermin (apd)基因家族3个新成员的特性鉴定及昆虫APD家族序列特征的分析 |
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| 引用本文： | 孙亮先,黄周英,郑华军,葛清秀,龚丽萍,陈怀宇.蜜蜂表皮蛋白apidermin (apd)基因家族3个新成员的特性鉴定及昆虫APD家族序列特征的分析[J].昆虫学报,2012,55(1):12-23. |
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| 作者姓名： | 孙亮先 黄周英 郑华军 葛清秀 龚丽萍 陈怀宇 |
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| 作者单位： | 泉州师范学院， 福建省高校分子生物学与药物化学重点实验室， 福建泉州 362000 |
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| 基金项目： | 国家自然科学基金项目，福建省高校服务海西建设重点项目，福建省教育厅省属高校项目 |
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| 摘 要： | Apidermin (APD)蛋白家族是一个新的昆虫结构性表皮蛋白家族。本研究结合生物信息学和RT-PCR扩增， 对意大利蜜蜂Apis mellifera ligustica（简称“意蜂”）的apd-1-like, apd-3-like和中华蜜蜂Apis cerena cerena（简称“中蜂”）的apd-2 等3个新的apd基因的结构特征和表达进行了分析， 并分析了昆虫APD蛋白家族的序列特征。结果显示， 在西方蜜蜂Apis mellifera（简称“西蜂”）中， apd基因家族的6个成员串联排列在基因组序列第4号连锁群上， 它们在A. m. ligustica雄蜂头部中的转录水平差异明显， 且其启动子序列所含顺式元件也不同。中蜂apd-2和意蜂apd-1-like都含有3个外显子和2个内含子， 而意蜂apd-3-like则由4个外显子和3个内含子组成。蛋白序列分析结果显示， 目前已知的10条APD蛋白序列N末端均具有相似的信号肽序列， 其成熟蛋白分子量为6.0~37.0 kD， pI为6.2~10.8。其中西蜂的APD1-3、APD-like和东方蜜蜂Apis cerena的APD-2等5条较短的多肽中疏水氨基酸残基达52%~67%， 且Ala含量最为丰富（占25%~34%）； 而丽蝇蛹集金小蜂Nasonia vitripennis的APD 1-3和西蜂APD-1-like， APD-3-like等另外5条APD多肽富含Gly（21%~30%）， 其序列中疏水氨基酸残基含量为35%~41%。多肽序列多重比对和系统进化分析结果显示， APD家族可划分为2个亚家族。亚家族Ⅰ含有西蜂APD 1-3和东方蜜蜂APD-2等4条较短的多肽序列， 其N末端为一个长33 aa的保守基序； 亚家族Ⅱ由另外6条相对较长的多肽序列组成， 其N末端保守基序长50 aa, C末端保守基序长16 aa。本文所描述的APD蛋白家族序列特征有助于以后从其他昆虫中鉴定新的apd基因。
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| 关 键 词： | 西方蜜蜂 东方蜜蜂 表皮蛋白 apidermin基因 APD家族 序列分析 生物信息学分析 |
Characterization of three new members of the apidermin (apd) gene family from honeybees and sequence analysis of the insect APD family |
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| SUN Liang-Xian
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HUANG Zhou-Ying
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ZHENG Hua-Jun
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GE Qing-Xiu
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GONG Li-Ping
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CHEN Huai-Yu.Characterization of three new members of the apidermin (apd) gene family from honeybees and sequence analysis of the insect APD family[J].Acta Entomologica Sinica,2012,55(1):12-23. |
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| Authors: | SUN Liang-Xian HUANG Zhou-Ying ZHENG Hua-Jun GE Qing-Xiu GONG Li-Ping CHEN Huai-Yu |
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| Affiliation: | The Higher Educational Key Laboratory for Molecular Biology and Pharmacology of Fujian Province, Quanzhou Normal University, Quanzhou, Fujian 362000, China |
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| Abstract: | The apidermin(APD) protein family is a novel structural cuticular protein family of insects.To gain a better understanding of this protein family,by using bioinformatics and RT-PCR amplification,we identified three novel apd genes(apd-1-like,apd-3-like from Apis mellifera ligustica,and apd-2 from Apis cerana cerana) and investigated their structural features,and then we revealed the characteristic motifs of the APD family.The results showed that a cluster of six apd genes were tandemly arranged on chromosome 4 of Apis mellifera.These apd genes were differentially expressed in drone head of A.m.ligustica,and their expression pattern was consistent with that of the cis-acting elements in their promoter sequences.The genomic DNA of apd-2 from A.c.cerana and apd-1-like from A.m.ligustica contain three exons and two introns,while that of apd-3-like from A.m.ligustica contain four exons and three introns.As the deduced proteins of the apd genes were analyzed,it was found that the presently available 10 APDs possess similar N-terminal signal peptide sequences.The mature APD proteins ranged in size from 6.0 to 37.0 kD,and their pI ranged from 6.2 to 10.8.Intriguingly,five small APD proteins,including APD-2 from A.cerana,and APD 1-3 and APD-like from A.mellifera,were rich in hydrophobic amino acid residues(52%-67%) with Ala being the most abundant(25%-34%).However,the other five larger APDs,including APD 1-3 from Nasonia vitripennis,and APD-1-like and APD-3-like from A.mellifera,were Gly-rich(21%-30%) proteins with hydrophobic amino acid residues constituting 35%-41% of their amino acid sequences.Multiple alignment and phylogenetic analysis revealed that the APD protein family could be classified into two subfamilies.Subfamily Ⅰ,which contains four low complexity sequences(APD 1-3 from A.mellifera and APD-2 from A.cerana),was characterized by a 33-aa N-terminal conserved motif.The other six larger APD proteins were classified as subfamilyⅡ,which was featured by a 50-aa N-terminal motif and a 16-aa C-terminal motif.The diagnostic features of APD protein family described here will be very helpful for identifying novel apd genes from arthropods. |
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| Keywords: | Apis mellifera Apis cerana cuticular protein apidermin gene APD family sequence analysis bioinformatical analysis |
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