亚洲玉米螟免疫凝集素基因的克隆、序列分析及组织表达
Molecular cloning, sequence analysis and tissue expression of immulectin gene from the Asian corn borer, Ostrinia furnacalis (Lepidoptera: Pyralidae)
刘 佳1, 2** 刘 洋1, 3 赵华福1 张文庆1 胡 建1***
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DOI：
作者单位：1. 中山大学生命科学学院，有害生物控制与资源利用国家重点实验室，广州 510275；2. 昆山科腾生物科技有限公司，苏州 215300；3. 苏州卓凯生物技术有限公司，苏州 215300
中文关键词：免疫凝集素，克隆，序列分析，组织表达，亚洲玉米螟
英文关键词： immulectin, clone, sequence analysis, tissue expression, Ostrinia furnacalis
中文摘要：
C型凝集素作为模式识别分子可以识别部分脂多糖（LPS），进而参与昆虫细胞的防御反应。本文通过RT-PCR和 3′/5′RACE技术从亚洲玉米螟Ostrinia furnacalis 5龄幼虫血细胞中克隆得到免疫凝集素基因（OfIML）。OfIML mRNA全长为1 241 bp，其中开放读码框（ORF）为924 bp，编码307个氨基酸（aa），分子量约为34.65 ku。与其它昆虫的C型凝集素比对分析结果显示，OfIML属于鳞翅目免疫凝集素，并且含有一个独特的结构特征，即一前一后2个糖识别域，氨基末端（CRD1，aa#1-135）和羧基末端（CRD2，aa#136-287）。RT-PCR检测OfIML在幼虫组织中的分布结果表明，其在血细胞、表皮、脂肪体、中肠、马氏管和气管中都有表达。OfIML GenBank 登录号为ABZ81710。OfIML 是一种昆虫免疫凝集素，含有2个糖识别域，根据其分子结构及在组织分布中的结果显示可能在亚洲玉米螟的免疫反应中起重要作用。
英文摘要：
C-type lectin, a kind of pattern recognition molecule, recognizes the lipid A portion of lipopolysaccharide (LPS) and participates the cellular defense reaction in insects. In this paper, an immulectin (OfIML), C-type lectin with two carbohydrate-recognition domains, was cloned by reverse-transcription PCR (RT-PCR) and 3′/5′RACE from the larval hemocytes of corn borer, Ostrinia furnacalis. The cDNA of OfIML is 1 241 base pairs in length, and contains an open reading frame (ORF) of 924 nucleotides, which encoding a protein of 307 amino acids with a predicted molecularmass of approximately 34.65 ku. Alignment of OfIML with C-type lectins of other insects indicates that OfIML is a member of Lepidoptera immulectins. OfIML possesses two carbohydrate-recognition domains (CRDs), an amino-terminal domain, CRD1 (residues 1–135), and a carboxyl-terminal domain, CRD2 (residues 136–287). RT-PCR analyses shows that OfIML is expressed in cuticle, fat body, midgut, tracheae, and malpighian tubules, especially in hemocytes. The cDNA sequence has been deposited with GenBank under accession No. ABZ81710. OfIML is a kind of insect immunlectin, which contains two carbohydrate-recognition domains, and it possibly plays an important role during the immune reaction of O. furnaclis depending on its molecular structure and expression in tissues.