杂色鲍AP-1的克隆及在发育和弧菌感染后的表达分析  

Molecular Cloning and Expression of Activator Protein-1 in Small Abalone Haliotis diversicolor during Development and Bacterial Challenge

作　　者：王国栋[1,2] 和四梅[1,2] 张丽莉[1,2] 王艺磊[1,2] 

机构地区：[1]集美大学水产学院,农业部东海海水健康养殖重点实验室,福建厦门361021 [2]集美大学水产生物技术研究所,福建厦门361021

出　　处：《水产科学》2016年第3期227-233,共7页Fisheries Science

基　　金：国家自然科学基金资助项目(41006105;41176152);福建省自然科学基金资助项目(2015J01142);福建省高校杰出青年科研人才项目(JA12183)

摘　　要：激活蛋白1(AP-1)是具有亮氨酸拉链功能域的转录因子,参与了发育和免疫等多个生物学过程。为研究该基因在杂色鲍发育和免疫中的作用,本研究克隆了杂色鲍AP-1基因的全长cDNA(命名为HdAP-1),并分析了HdAP-1在各组织、各发育时期以及副溶血弧菌刺激后的表达特征。结果表明,HdAP-1cDNA全长为1482bp,5′非编码区域(5′UTR)为133bp,3′UTR为401bp,开放阅读框为948bp。该基因编码蛋白预测分子量为34.83ku,等电点为9.43,具有典型AP-1蛋白家族的Jun转录因子功能域和bZIP功能域。HdAP-1在所检测7个组织中均有表达,其中血淋巴表达量最高,鳃、肾、肠和黏液腺的表达量次之,肝胰腺和外套膜表达量最低。自受精卵至囊胚期HdAP-1的表达量显著低于原肠胚(P<0.05),由原肠胚到担轮幼虫该基因表达量继续上升,担轮幼虫至匍匐幼虫期均维持较高的表达水平,变态后在稚鲍中的表达量下降。在副溶血弧菌感染24h后,HdAP-1表达量显著上升(P<0.05)。其他时段该基因的表达量与对照组相比差异不显著(P>0.05)。Activator protein-1 （AP-1） is a transcription factor, which is characterized by highly conserved dimeric leucine zipper DNA-binding domain, and participates in development and immune processes. In or der to evaluate the effect of AP 1 gene in development and immune response, the full cDNA of AP-1 gene （HdAP-1） was clored in small abalone Haliotis diversicolor and challenged by Vibrio parahaemolyticus and expressed during development of the abalone by real time PCR. The full cDNA of HdAP-1 was 1482 bp, and 5＇/3＇untranslated region had 133/401 bp, with a 948 bp open reading frame. The calculated mo- lecular mass of encoded protein was 34.83 ku and the predicted pI 9.43. HdAP 1 contained Jun and bZIP domains, a highly conserved domains of AP-1 protein family. HdAP-1 was detected in all examined tis- sues. There was the maxmal expression level in hemolymph, middle expression level in gill/kidney/diges- tive tract/mucus gland, the minimal level in hepatopancreas/mantle. There was a low expression level from fertilized eggs to blastula stage, significantly lower than in gastrula stage （P〈0.05）. The expression of HdAP-1 continuously increased from gastrula stage to trochophore stage, with a high level from tro- chophore to creeping larva stage, and low level after metamorphosis. There was significant up-regulation of HdAP-1 24 h after V. parahaemolyticus challenge （P〈0.05）, without significant difference between control and bacterial challenge in other phase （P〉0.05）.

关 键 词：杂色鲍 激活蛋白(AP-1) 基因表达 发育 细菌感染 

分 类 号：S917[农业科学—水产科学]