水生生物学报2018，Vol.42Issue(1)：68-76,9.DOI:10.7541/2018.009
金鱼胰蛋白酶原基因的克隆及镉和过氧化氢暴露对其基因表达的影响
MOLECULAR CLONING AND CHARACTERIZATION OF TRYPSINOGEN GENE IN GOLDFISH (CARASSIUS AURATUS) AND ITS EXPRESSION PROFILES IN RE-SPONSE TO CADMIUM AND OXIDATIVE STRESS
摘要
Abstract
Trypsin is a serine protease that plays a major role in protein digestion. It is synthesized and secreted by pancreas as a prepro enzyme, trypsinogen. In the present study, a full-length cDNA of goldfish trypsinogen (gfTryp) was successfully cloned from the hepatopancreas by rapid amplification of cDNA ends technique. The obtained gfTryp cDNA was 864 bp long with a 21 bp 5′-untranslated region (UTR), a 114 bp 3′-UTR containing the consensus polyadenylation signal AATAAA, and a 729 bp open reading frame encod-ing a protein of 242 amino acid residues. Sequence alignment showed that the gfTryp possessed all the chara-cteristic features of trypsin family, suggesting its conserved function in protein digestion. Its mRNA expres-sion was observed in all tissues examined, and the relatively higher levels were detected in hepatopancreas, intestine and fat. Hepatopancreatic gfTryp mRNA level decreased significantly after fasting for one week. Further periprandial expression analysis showed that gfTryp mRNA expression level in hepatopancreas dra-matically up-regulated after meal. Hepatopancreatic gfTryp mRNA expression level increased significantly with 0.5 and 5 μg/mL cadmium (3.2 and 4.7 fold, respectively), and decreased with cadmium concentration up to 10 μg/mL. Strikingly, H2O2 exposure significantly decreased gfTryp mRNA expression in hepatopan-creas at 3h, 6h, 12h and 24h. Our study provides the evidence that nutritional status, cadmium and oxidative stress can regulate the trypsinogen expression at the transcript level, and sheds new light on the physiological expression of trypsinogen in fish.关键词
金鱼/胰蛋白酶原/营养状态/镉/氧化应激/mRNA表达Key words
Carassius auratus/Trypsinogen/Nutritional status/Cadmium/Oxidative stress/mRNA expression分类
农业科技引用本文复制引用
陈文波,闫芳芳,秦少宗,娄本杰..金鱼胰蛋白酶原基因的克隆及镉和过氧化氢暴露对其基因表达的影响[J].水生生物学报,2018,42(1):68-76,9.基金项目
Supported by the National Natural Science Foundation of China (31101878) （31101878）
the Excellent Youth Foundation of Henan Poly-technic University (J2014-02) （J2014-02）
the Youth Backbone Teacher Foundation of Henan Polytechnic University (672105/126) （672105/126）
the Key Science Research Project in University of Henan Province (16A240001) （16A240001）
