东亚飞蝗中肠几丁质酶基因的克隆、序列分析及组织定位  

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

作　　者：张八生[1] 曹月青[1] 张伟[1] 殷幼平[1] 王中康[1] 彭国雄[1] 夏玉先[1] 

机构地区：[1]重庆大学生物工程学院基因工程研究中心

出　　处：《昆虫学报》2007年第6期555-559,共5页Acta Entomologica Sinica

基　　金：国家"863"项目(2004AA246050);教育部重点项目(0317)

摘　　要：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1604bp,其中可读框1452bp,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。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.

关 键 词：东亚飞蝗 几丁质酶基因 中肠 半定量RT-PCR 

分 类 号：Q966[生物学—昆虫学]