拟穴青蟹丝裂原活化蛋白激酶激酶(MAPKK)基因的克隆与表达分析

Abstract

采用QrT-PCr、rACE等方法,获得了拟穴青蟹丝裂原活化蛋白激酶激酶(MAPkk)基因CdnA全长序列。该基因全长1 558 bP,开放阅读框长度为1 224 bP,编码407个氨基酸残基。同源分析显示,该基因编码的蛋白与昆虫的相似性高达70%,推测MAPkk基因在节肢动物具有较高的保守性。经荧光定量PCr检测,MAPkk基因在拟穴青蟹多个组织中有表达,且在脑神经节和卵巢中表达量较高。在拟穴青蟹卵巢发育过程中,MAPkk基因在卵巢发育期(Ⅲ期)表达量最高,发育期为卵母细胞快速生长期,推测MAPkk具有促进卵母细胞快速生长的作用。

In this paper,the MAPKK( mitogen-activated protein kinase kinase) was isolated from the mud crab,Scylla paramamosain using RT-PCR and RACE methods.The obtained full-length cDNA of MAPKK w as 1 558 bp w ith an open reading frame of 1 224 bp encoding a putative peptide of 407 amino acids.By alignment,the amino acid sequence of S.paramamosain MAPKK show ed high homology w ith those of some other animals.It suggested MAPKK w as highly conservative.Real-time PCR show ed that the MAPKK gene w as expressed in various tissues,and highly expressed in brain ganglion and ovary.The MAPKK mRNA profiles during ovarian development indicated that the expression of MAPKK w as significantly high at developing stage.We inferred that MAPKK might play a stimulative role in the ovarian development of the mud crab.