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).