白甲乌鳢酪氨酸酶(TYR)基因的克隆,表达及其分子特征分析
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- 英文论文题名：Molecular cloning, characterization and expression analysis of TYR gene in Opniocepnalus argus var Kimnra
- 论文作者：周爱国 ; 范兰芬 ; 王桢璐 ; 叶峭 ; 姜文钊 ; 邹记兴
- 英文论文作者：ZHOU Aiguo ; FAN Lanfen ; WANG Zhenlu ; YE Qiao ; JIANG Wenzhao ; ZOU Jixing ; College of Marine Sciences ; South China Agricultural University ; Qingyuan North River Fishery Science Institute
- 年：2016
- 作者机构：华南农业大学海洋学院;清远市北江水产科学研究所;
- 论文关键词：白甲乌鳢 ; TYR ; 克隆 ; 表达 ; 分子特征 ; 亚细胞定位
- 英文论文关键词：Opniocepnalus argus var Kimnra ; TYR gene ; cloning ; expression ; molecular characteristics ; subcellular localization
- 会议召开时间：2016-11-02
- 会议录名称：2016年中国水产学会学术年会论文摘要集
- 语种：中文
- 分类号：S917.4
- 学会代码：OGSB
- 会议名称：2016年中国水产学会学术年会
- 会议地点：中国四川成都
- 主办单位：中国水产学会、四川省水产学会
- 学会名称：中国水产学会
- 页数：1
- 文件大小：560k
- 原文格式：D
- 会议级别：全国
摘要
为了初步探索白甲乌鳢的白化特征机理,本实验利用RACE技术克隆了白甲乌鳢和普通乌鳢TYR基因cDNA全长,两者序列一致。同源比对发现,与硬骨鱼类相似性较高,而与细菌类相似度较低。相对实时荧光定量PCR结果显示TYR基因在白甲乌鳢和乌鳢9大组织(皮肤、眼睛、肌肉、心脏、肝脏、脾脏、肾脏、鳃和肠道)中均有表达。其中,白甲乌鳢眼睛组织表达量最高,而乌鳢皮肤组织的表达量最高。对比相同组织发现,乌鳢皮肤组织TYR基因表达量极显著高于白甲乌鳢(18.66倍)。细胞免疫荧光观察亚细胞定位于内质网、高尔基体、细胞质膜等部位,与预测结果一致。可见,TYR基因可能作为参与乌鳢皮肤黑色素细胞发育、分化和功能调节的重要因子,其低表达是皮肤出现白化特征的关键因素之一。
In order to explore the mechanism of Opniocepnalus argus var Kimnra, the full-length cDNA of Opniocepnalus argus var Kimnra and Channa argus TYR gene were cloned and characterized, BLAST analysis showed that the sequences between them are consistent.Homology analysis indicated that it has a highest similarity with teleost, but a low similarity with bacterias. Relative quantitative real-time PCR(qRT-PCR) revealed tissue specific TYR expression. The highest expression is in eyes of Opniocepnalus argus var Kimnra, while in skin of Channa argus. Compare the same tissues, the expression in skin of Channa argus is significantly higher than Opniocepnalus argus var Kimnra. Then the immunofluorescence observation of subcellular localization was mainly distributed in the endoplasmic reticulum and Golgi membrane,cytoplasm, consistented with the prediction results. From these results we can know that the TYR may be involved in the development, differentiation and functional regulation of the skin melanoma cells. Its low expression is a one of the key factor leading to the skin albinotic characteristics.
In order to explore the mechanism of Opniocepnalus argus var Kimnra, the full-length cDNA of Opniocepnalus argus var Kimnra and Channa argus TYR gene were cloned and characterized, BLAST analysis showed that the sequences between them are consistent.Homology analysis indicated that it has a highest similarity with teleost, but a low similarity with bacterias. Relative quantitative real-time PCR(qRT-PCR) revealed tissue specific TYR expression. The highest expression is in eyes of Opniocepnalus argus var Kimnra, while in skin of Channa argus. Compare the same tissues, the expression in skin of Channa argus is significantly higher than Opniocepnalus argus var Kimnra. Then the immunofluorescence observation of subcellular localization was mainly distributed in the endoplasmic reticulum and Golgi membrane,cytoplasm, consistented with the prediction results. From these results we can know that the TYR may be involved in the development, differentiation and functional regulation of the skin melanoma cells. Its low expression is a one of the key factor leading to the skin albinotic characteristics.
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