东亚飞蝗中肠几丁质酶基因的克隆、序列分析及组织定位

通过RACE方法，克隆了东亚飞蝗Locusta migratoria manilensis (Meyen)几丁质酶基因 (LmChi)cDNA全序列 (GenBank 登录号：EF092841)。获得的cDNA全长1 604 bp，其中可读框1 452 bp, 编码483个氨基酸。推测其氨基酸序列与18家族昆虫几丁质酶有较高的相似性。与其他几丁质酶一样，东亚飞蝗几丁质酶序列也包含一个信号肽、一个几丁质酶活性位点、一个碳端丝氨酸富集区和一个几丁质结合域。半定量RT-PCR研究表明，LmChi基因只在东亚飞蝗不同发育阶段的中肠组织中表达，而在东亚飞蝗体壁、前肠和后肠均没有发现LmChi基因的转录。

Cloning, sequence analysis and tissue expression of chitinase gene from the midgut of Locusta migratoria manilensis (Meyen)

The full cDNA of chitinase gene (LmChi) from the midgut of Locusta migratoria manilensis (Meyen) was cloned by means of 3′-RACE and 5′-RACE. The results showed that the cDNA was 1 604 bp in length and contained an open reading frame (ORF) of 1 452 bp (GenBank accession number: EF092841), which encoded 483 amino acids. The deduced amino acid sequence, which was composed of a signal peptide, a chitinase active site, a C-terminal serine-rich region and a domain normal present in chitinase, showed a high similarity with other insect chitinases from family 18.Semi-quantitative RT-PCR indicated that LmChi expressed only in the midgut, but not in the integument, foregut and hindgut of L. migratoria manilensis.