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黄颡鱼HSC70基因及其组织表达分析
  • ISSN号:1000-3207
  • 期刊名称:《水生生物学报》
  • 时间:0
  • 分类:Q781[生物学—分子生物学]
  • 作者机构:[1]暨南大学水生生物研究所,广州510632, [2]水产科学重庆市市级重点实验室、三峡库区生态环境教育部重点实验室,重庆400715
  • 相关基金:国家自然科学基金项目(40576056);广东省自然科学基金项目(04300664,07300378);中国博士后科学基金一等资助金(20070420144)资助
中文摘要:

热休克蛋白70(HSP70)与生物体的抗胁迫能力密切相关。本文采用RACE(Rapid amplification of eDNA ends)技术,从黄颡鱼Pelteobagrus fulvidraco克隆到一种组成型热休克蛋白(HSC70)基因及其cDNA。该cDNA全长2245bp,包括5’非编码区82bp,3’非编码区225bp,开放阅读框(ORF)1938bp,编码645个氨基酸组成的蛋白质。黄颡鱼HSC70基因含有8个内含子,与人、鼠、虹鳟和花斑溪鳝的HSC70基因内含子数目相同,位置相似。其中,最长内含子(873bp)位于5’端非编码区,其余内含子(长度在80-251bp之间不等)均在编码区以内。黄颡鱼HSCTO基因编码的氨基酸序列与南方鲶的相似度最高,达96.13%,与欧洲银鲫和团头鲂的相似度分别为94.45%和94.14%。RT—PCR检测显示,正常情况下黄颡鱼HSC70在血细胞、心脏、肝、头肾、脾、鳃、肌肉和脑中均有表达,但表达量在鳃中最高,肌肉中最低;统计结果显示,热激后HSC70在血细胞、肝、头肾和脑中的表达量显著上升(P〈0.05),而在其余组织中热激前后的表达差异不显著(P〉0.05)。

英文摘要:

Heat shock protein 70 (HSP70s) act as a role of chaperone and play a key function in cytoprotection and cytorepair, including protein assembly, correct folding, and membrane translocation, it also enhance the organisms' immunity and enduration to stressors. Yellow catfish (Pelteobagrus fulvidraco) is an important cultured species in China. In order to illuminate molecular mechanism of the HSP70 family members in the catfish against stressors and diseases, it is necessary to clone the gene and eDNA sequence of HSP70 family members in the first instance. Therefore, the gene and its eDNA of a HSP70 family member were cloned in yellow catfish, and mRNA expression of the gene was studied in various tissues and organs of the catfish under heat-treated or unstressed condition. A full length eDNA of 2245 bp was cloned in the gill of yellow catfish with RACE (rapid amplification of eDNA ends) technique. The eDNA contained an open reading frame (ORF) of 1938 bp, 5' untranslated region of 82 bp and 3' untranslated region of 225 bp. The deduced 645 amino acid sequence contained HSP70s' characteristic motifs (Fig. 2), and it indicated that the eDNA belonged to the family of heat shock protein 70. Carried out alignment with other organisms' HSC70 amino acid sequences, the deduced amino acid sequence from the eDNA showed the highest similarity (96.13%) with HSC70 amino acid sequence of Southern catfish (Silurus meridionalis). The phylogenetic tree ( Fig. 3 ) showed that 70 kD heat shock protein of yellow catfish clustered together with other vertebrates' HSC70s but not HSP70s. What mentioned above suggested that the sequence we cloned was a kind of heat shock cognate 70 (HSC70). Furthermore, its constitutive expression in unstressed tissue cells by RT-PCR detection ( Fig. 4) confirmed that the sequence we cloned was HSC70 eDNA. Subsequently, the catfish HSC70 gene was cloned by PCR amplification in the fish genome DNA. Eight introns were found in the HSC70 gene, and the longe

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期刊信息
  • 《水生生物学报》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学院
  • 主办单位:中科院水生所
  • 主编:桂建芳
  • 地址:武昌东湖南路7号中科院水生所
  • 邮编:430072
  • 邮箱:Acta@ihb.ac.cn
  • 电话:027-68780701
  • 国际标准刊号:ISSN:1000-3207
  • 国内统一刊号:ISSN:42-1230/Q
  • 邮发代号:82-329
  • 获奖情况:
  • 湖北省十佳重点期刊,中国农学会中国水产学会优秀科技期刊一等奖,中国科学院优秀期刊三等奖,中国期刊方阵“双效”期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,美国生物科学数据库,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:21674