小地老虎β-N-乙酰葡萄糖胺糖苷酶基因cDNA序列的克隆及mRNA组织特异性表达
cDNA cloning and mRNA expression of β-N-acetylglucosaminidase indifferent tissues of Agrotis ipsilon Hufngel
高艳玲 吴丽梅 郭博智 樊东
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作者单位：东北农业大学农学院哈尔滨150030
中文关键词： 小地老虎，β-N-乙酰葡萄糖胺糖苷酶，分子克隆，组织表达
英文关键词： Agrotis ipsilon，β-N-acetylglucosaminidase，cloning，mRNA expression
中文摘要：β-N-乙酰葡萄糖胺糖苷酶是昆虫几丁质代谢中一种关键酶，在昆虫的多种生理活动中发挥重要作用。本文以小地老虎Agrotis ipsilon Hufngel 预蛹期幼虫为材料提取总RNA，利用RT-PCR 和RACE 技术，扩增得到其β-N-乙酰葡萄糖胺糖苷酶基因的cDNA 序列。该序列含有2 701 个碱基，包括一个1 788 个碱基的开放阅读框，编码一个含595 个氨基酸的蛋白，分子量约为68. 3 ku，等电点为5. 48。推导得到的氨基酸序列与其他昆虫，尤其是鳞翅目昆虫的β-N-乙酰葡萄糖胺糖苷酶高度同源。基因cDNA 序列已经登录GenBank 并获得登录号GU985280。该基因在小地老虎取食期和预蛹期的马氏管、中肠、脂肪体、体壁和去中肠虫体中均含有mRNA 水平的表达。
英文摘要：β-N-acetylglucosaminidase is an enzyme that can degrade chitin in insects． Total RNA was isolated from theprepupal stage of Agrotis ipsilon Hufngel． The cDNA sequence was cloned by RT-PCR and rapid amplification of cDNAends(RACE) ． The cDNA，2 701 base pairs in length，contained an open reading frame of 1 788 base pairs coding for a polypeptide of 595 amino acid residues with a predicted molecular weight of 68. 3 ku and pI 5. 48． The predicted proteinshared extensive similarities with those from other insects，expecially Lepidopteran insects． The cDNA sequence has beendeposited in GenBank with the accession No． GU985280． RT-PCR results indicate that β-N-acetylglucosaminidase mRNA
was expressed in the malpighian tubules，midgut，fat bodies and integument．