Cheng Peizhou, Liu Xiao, Zhang Guofan, He Jianguo
Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, Shandong, PR China.
Fish Shellfish Immunol. 2007 Jan-Feb;22(1-2):77-87. doi: 10.1016/j.fsi.2006.03.014. Epub 2006 Apr 4.
Heat shock protein 70 (HSP70), the primary member of HSPs that are responsive of thermal stress, is found in all multicellular organisms and functions mostly as molecular chaperon. The inducible HSP70 cDNA cloned from Pacific abalone (Haliotis discus hannai) using rapid amplification of cDNA ends (RACE), was highly homologous to other HSP70 genes. The full-length cDNA of the Pacific abalone HSP70 was 2631bp, consisting of a 5'-terminal untranslated region (UTR) of 90bp, a 3'-terminal UTR of 573bp with a canonical polyadenylation signal sequence AATAAA and a poly (A) tail, and an open reading frame of 1968bp. The HSP70 cDNA encoded a polypeptide of 655 amino acids with an ATPase domain of 382 amino acids, the substrate peptide binding domain of 161 amino acids and a C-terminus domain of 112 amino acids. The temporal expression of HSP70 was measured by semi-quantitative RT-PCR after heat shock and bacterial challenge. Challenge of Pacific abalone with heat shock or the pathogenic bacteria Vibrio anguillarum resulted in a dramatic increase in the expression of HSP70 mRNA level in muscle, followed by a recovery to normal level after 96h. Unlike the muscle, the levels of HSP70 expression in gills reached the top at 12h and maintained a relatively high level compared with the control after thermal and bacterial challenge. The upregulated mRNA expression of HSP70 in the abalone following heat shock and infection response indicates that the HSP70 gene is inducible and involved in immune response.
热休克蛋白70（HSP70）是热应激反应性热休克蛋白（HSPs）的主要成员，存在于所有多细胞生物中，主要发挥分子伴侣的功能。利用cDNA末端快速扩增技术（RACE）从太平洋鲍鱼（皱纹盘鲍）中克隆得到的可诱导型HSP70 cDNA，与其他HSP70基因高度同源。太平洋鲍鱼HSP70的全长cDNA为2631bp，由一个90bp的5'-末端非翻译区（UTR）、一个573bp的3'-末端UTR（带有典型的多聚腺苷酸化信号序列AATAAA和一个poly（A）尾）以及一个1968bp的开放阅读框组成。HSP70 cDNA编码一个由655个氨基酸组成的多肽，其中包含一个由382个氨基酸组成的ATP酶结构域、一个由161个氨基酸组成的底物肽结合结构域以及一个由112个氨基酸组成的C末端结构域。热休克和细菌攻击后，通过半定量RT-PCR检测HSP70的时序表达。对太平洋鲍鱼进行热休克或致病性鳗弧菌攻击后，肌肉中HSP70 mRNA水平的表达显著增加，96小时后恢复到正常水平。与肌肉不同，鳃中HSP70的表达水平在12小时达到峰值，在热应激和细菌攻击后与对照组相比维持在相对较高的水平。热休克和感染反应后鲍鱼中HSP70的mRNA表达上调，表明HSP70基因是可诱导的，并且参与免疫反应。
