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引用本文:王志刚,刘士力,李贤露,吕卓云,毛丽盈,唐超然,郑荣泉.翘嘴红鲌(Erythroculter ilishaeformis)果糖-1,6-二磷酸醛缩酶(ALDO-C)基因定位、克隆及表达分析.海洋与湖沼,2019,50(5):1138-1145.
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翘嘴红鲌(Erythroculter ilishaeformis)果糖-1,6-二磷酸醛缩酶(ALDO-C)基因定位、克隆及表达分析
王志刚1, 刘士力2, 李贤露1, 吕卓云1, 毛丽盈1, 唐超然1, 郑荣泉1,3,4
1.浙江师范大学化学与生命科学学院 金华 321004;2.浙江省淡水水产研究所 农业农村部淡水渔业健康养殖重点实验室/浙江省淡水水产遗传育种重点实验室 湖州 313001;3.浙江省野生动物生物技术与保护利用重点实验室 金华 321004;4.浙江师范大学行知学院 金华 321004
摘要:
翘嘴红鲌(Erythroculter ilishaeformis)是大型淡水鱼类鳊鲌亚科(Abramidinae)中最大的一种鱼,具有较高的经济价值。但其饲料转化率及抗病性研究相对较少,相关基因信息缺乏。本研究以翘嘴红鲌为对象,利用RACE技术克隆翘嘴红鲌果糖-1,6-二磷酸醛缩酶(ALDO-C)基因,该基因cDNA全长1945bp,其中ORF区975bp,编码325个氨基酸,5'非编码区933bp,3'非编码区37bp。通过实时荧光定量PCR检测了ALDO-C在不同组织中相对表达水平。发现ALDO-C基因在翘嘴红鲌的肾、肝、肌肉、性腺中均有表达,且在肾中表达量最高,显著高于其他组织。同时采用石蜡切片、H.E染色和原位杂交染色,观察分析翘嘴红鲌的肾组织显微结构和ALDO-C基因的表达定位,结果表明,ALDO-C在鱼类肾单位和集合管结构、调节肾组织水平衡及抗病性方面有重要作用。可以考虑将翘嘴红鲌醛缩酶C基因作为生长发育及抗病性相关的候选基因,用于翘嘴红鲌鱼的分子辅助育种,以期为今后的研究提供理论基础。
关键词:  翘嘴红鲌  ALDO-C    原位杂交
DOI:10.11693/hyhz20190300049
分类号:Q786;S917
基金项目:浙江省农业(水产)新品种选育重大科技专项,2016C02055-1号。
LOCATION, CLONING AND EXPRESSION ANALYSIS OF ALDO-C GENE IN ERYTHROCULTER ILISHAEFORMIS
WANG Zhi-Gang1, LIU Shi-Li2, LI Xian-Lu1, Lü Zhuo-Yun1, MAO Li-Ying1, TANG Chao-Ran1, ZHENG Rong-Quan1,3,4
1.College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China;2.Agriculture Ministry Key Laboratory of Healthy Freshwater Aquaculture/Key Laboratory of Freshwater Aquatic Animal Genetic and Breeding of Zhejiang Province, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China;3.Key Lab of Wildlife Biotechnology and Conservation and Utilization of Zhejiang Province, Jinhua 321004, China;4.Xingzhi College, Zhejiang Normal University, Jinhua 321004, China
Abstract:
Erythroculter ilishaeformis, or common skygazer, is a high-economic-valued large freshwater fish species of Subfamily Paralichthys. However, studies on the feed conversion rate and disease resistance for the culture are scarce, and the understanding of the related gene information remains poor. We cloned fructose-1,6-diphosphate aldolase (ALDO-C) gene by RACE (rapid-amplification of cDNA ends) technique. The full length of the gene was 1945bp, including 975bp in ORF (open reading frame) region, 325 amino acids, 933bp in 5'non-coding region, and 37bp in 3' non-coding region. The relative expression levels of ALDO-C in different tissues were detected by real-time fluorescence quantitative PCR (polymerase chain reaction). It was found that ALDO-C gene was expressed in kidney, liver, muscle, and gonad, and the highest expression was found in kidney, being significantly higher than those in other tissues are. At the same time, paraffin section, H&E (hematoxylin and eosin) staining, and in situ hybridization staining were used to observe and analyze the microscopic structure of kidney tissue and the expression and location of ALDO-C gene. The results show that ALDO-C played an important role in structuring the nephron and collecting duct, regulating renal tissue water balance, and resisting against disease. Therefore, the aldolase C gene can be considered as a candidate gene relating to the growth, development, and disease resistance for molecular-assisted breeding of the fish. This study provided a theoretical basis for future research in this regard.
Key words:  Erythroculter ilishaeformis  ALDO-C  kidney  in situ hybridization
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