| 摘要: |
| 【目的】克隆草鱼杀菌通透性增加蛋白/脂多糖结合蛋白（Bactericidal permeability-increasing protein/LPS binding protein,BPI/LBP）基因的cDNA全长，分析其结构特征，研究其表达规律，为进一步探明其功能奠定基础。【方法】首先用同源克隆及快速扩增cDNA末端技术(RACE)，克隆草鱼BPI/LBP基因的cDNA全长，然后用生物信息学方法分析BPI/LBP基因的结构特征，最后用半定量RT-PCR检测BPI/LBP基因在草鱼不同组织（血、脑、眼睛、前肠、中肠、后肠、鳔、鳃、头肾、中肾、心脏、肝胰脏、肌肉、皮肤和脾脏）中的表达模式。【结果】草鱼BPI/LBP cDNA全长1 568 bp，编码473个氨基酸，其中包括18个氨基酸组成的信号肽、BPI1结构域（BPI/LBP/CETP N-terminal domain）和BPI2结构域（BPI/LBP/CETP C-terminal domain）。该蛋白的分子质量为51 551 u，等电点为8.69。氨基酸序列同源性分析结果显示，草鱼与鲤鱼BPI/LBP的同源性最高（90%），其次为虹鳟（73%）、斑点叉尾鮰（73%）、香鱼（72%）等。在系统进化树中，草鱼BPI/LBP首先与鲤科的鲤鱼聚为一类。通过半定量RT-PCR分析可知，草鱼BPI/LBP基因在被检测的15个组织中都有表达，其中在鳃中的表达量最高，其次为头肾、中肾等。【结论】成功克隆了草鱼BPI/LBP基因的cDNA全长，该基因在草鱼体内不同组织中广泛表达。 |
| 关键词: 草鱼 杀菌通透性增加蛋白/脂多糖结合蛋白 基因克隆 特征分析 |
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| 基金项目:国家自然科学基金项目(30871917)；教育部新世纪优秀人才支持计划项目(NCET-08-0466) |
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| Cloning and characterization of BPI/LBP gene from grass carp,Ctenopharyngodon idella |
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| Abstract: |
| 【Objective】The full-length cDNA sequence of bactericidal permeability-increasing protein/LPS binding protein (CiBPI/LBP) from grass carp,Ctenopharyngodon idella was cloned to study the characterization and expression profile and to lay a foundation for the functional studies.【Methods】The CiBPI/LBP cDNA was identified from grass carp gill by homologous cloning and rapid amplification cDNA ends (RACE).The structural characterization was analyzed by bioinformatics method.The CiBPI/LBP expression pattern was examined in different tissues (blood,brain,eye,foregut,midgut,hindgut,gas blander,gill,head kidney,trunk kidney,heart,liver,muscle,skin and spleen) by semi-quantitative RT-PCR.【Result】The CiBPI/LBP cDNA was 1 568 bp,encoding 473 amino acid (aa) residues,including signal peptide,BPI1 domain (BPI/LBP/CETP N-terminal domain) and BPI2 domain (BPI/LBP/CETP C-terminal domain).The molecular weight of the deduced protein was 51 551 u,and the isoelectric point 8.69.The amino acid sequence of CiBPI/LBP possessed 90%,73%,73%,and 72% identities with the BPI/LBPs of Cyprinus carpio,Oncorhynchus mykiss,Ictalurus punctatus and Plecoglossus altivelis altivelis respectively.CiBPI/LBP protein firstly clustered with BPI/LBP in Cyprinidae species,Cyprinus carpio in the phylogenetic analysis.CiBPI/LBP mRNA was detected in all the 15 tested tissues by semi-quantitative real-time RT-PCR,high in gill,head kidney and trunck kidney. 【Conclusion】The full-length CiBPI/LBP cDNA sequence was cloned successfully.CiBPI/LBP expression was ubiquitous. |
| Key words: Grass carp(Ctenopharyngodon idella) BPI/LBP gene cloning characterization |
