三疣梭子蟹fps克隆及其在卵巢发育中的表达分析  

Cloning of farnesyl diphosphate synthase(fps) from Portunus trituberculatus and its expression during the ovarian development

作　　者：朱韬[1] 朱冬发[1] 邱锡尔[1] 周彦琦 柳志业 谢熙[1] 

机构地区：[1]宁波大学海洋学院,宁波315211

出　　处：《生物学杂志》2016年第3期10-14,19,共6页Journal of Biology

基　　金：国家自然科学基金项目(41376152);浙江省自然科学基金项目(LY13C190006)资助

摘　　要：法尼基焦磷酸合成酶(FPS)是类异戊二烯生物合成途径中的关键酶,参与甲壳动物甲基法尼酯(MF)等萜类激素的合成。为研究FPS在甲壳动物卵巢发育中的调控作用,采用RT-PCR和cDNA末端快速扩增技术(RACE技术)克隆得到了三疣梭子蟹fps(Ptfps)的序列全长(Gen Bank登录号:KM013803)。结果显示:Ptfps cDNA全长为2 360 bp,开放阅读框长1 290 bp,编码429个氨基酸。生物信息学分析发现PtFPS具有7个异戊烯基转移酶的保守结构域,与已公布的内华达古白蚁FPS同源性最高(达到58%)。实时荧光定量PCR分析结果显示:fps主要在大颚器(MO)中表达,远高于其它组织的表达水平(P<0.01);在第1次卵巢发育过程中,fps的表达水平在Ⅳ期显著升高(P<0.05),之后略有下降;在第2次卵巢发育过程中,fps的表达水平在Ⅲ期显著升高(P<0.05),并在IV期达到最大。以上结果表明fps可能参与三疣梭子蟹卵巢发育的调控。Farnesyl diphosphate synthase（ FPS） is a key enzyme in isoprenoid biosynthesis,participates in the synthesis of methyl farnesoate（ MF） in the crustaceans. To study the regulatory role of FPS during ovarian development of the crustaceans,a full-length of fps（ Ptfps,Gene Bank accession number： KM013803） was cloned from Portunus trituberculatus by RT-PCR and rapid amplification of cDNA ends（ RACE）. The full-length of Ptfps was 2 360 bp and it contained an open reading frame（ ORF） of 1 290 bp encoding a protein of 429 amino acid residues. Bioinformatics analysis found that Pt FPS has seven prenyltransferase conserved regions,and exhibits the highest identity（ 58%） with FPS of Zootermopsis nevadensis. Quantitative real-time PCR analysis results showed that fps was mainly expressed in mandibular organ（ MO）,and the expression level was much higher than that in other tissues（ P 0. 01）. During the first ovarian development,the expression of fps significantly increased to the highest levels at stage Ⅳ（ P 0. 05）,and then slightly declined; During the second ovarian development,the expression of fps significantly increased to the highest level at stage Ⅲ（ P 0. 05）,and reached to the maximum at stage Ⅳ. The above results indicated that fps may closely related to the regulation of ovarian development in P. trituberculatus.

关 键 词：三疣梭子蟹 法尼基焦磷酸合成酶 克隆 卵巢发育 表达水平 

分 类 号：Q78[生物学—分子生物学]