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‐1 cDNA 全长为1482 bp ，5′非编码区域（5′UTR）为133 bp ，3′UTR为401 bp ，开放阅读框为948 bp。该基因编码蛋白预测分子量为34．83 ku ，等电点为9．43，具有典型A P‐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24 h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）。
Abstract：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 cloned 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) .
收稿日期: 2016-03-25
引用本文:
王国栋,和四梅,张丽莉,王艺磊. 杂色鲍 AP-1的克隆及在发育和弧菌感染后的表达分析[J]. 水产科学, 2016, 35(3): 227-233.
WANG Guodong,HE Simei,ZHANG Lili,WANG Yilei. Molecular Cloning and Expression of Activator Protein-1 in Small Abalone Haliotis diversicolor during Development and Bacterial Challenge. Fisheries Science, 2016, 35(3): 227-233.
