草鱼(Ctenopharyngodon idellus)NCCRP-1基因的克隆和原核表达

摘要/Abstract

摘要： 应用RACE PCR克隆了草鱼NCCRP-1 基因。结果表明，NCCRP-1 cDNA全长900 bp，开放阅读框为714 bp，编码237个氨基酸残基，预测分子量为27.3 kD，理论等电点为5.63。草鱼 NCCRP-1具有NCCRP-1蛋白家族保守的功能区域，与鲤鱼该基因(Cyprinus carpio L.)的相似性为 88%。将此基因定向插入原核表达载体pET-32a，构建重组质粒pET-NCCRP后转化BL21(DE3)进行诱导表达。SDS-PAGE分析显示在47.8 kD处出现特异性蛋白条带，Western blot检测表明草鱼NCCRP-1融合蛋白成功表达。

关键词: 草鱼, NCCRP-1, 克隆, 原核表达

Abstract: In the present study, grass carp(Ctenopharyngodon idellus)NCCRP-1 gene was cloned using RACE method. The full length cDNA of NCCRP-1 was 900 bp containing a 714 bp open reading frame(ORF), which encoded 237 amino acids with predicted molecular mass of 27.3 kD and a theoretical pI of 5.63. Amino acid alignment indicated that grass carp NCCRP-1 possessed consensus functional domians of NCCRP-1 family and shared 88% similarity with common carp NCCRP-1. The prokaryotic expression vector pET-NCCRP was constructed and then transformed into BL21(DE3). SDS-PAGE and Western blotting analysis indicated that the recombinant protein of grass carp NCCRP-1 was successfully expressed and molecular weight of expressed fusion protein was 47.8 kD.

Key words: Ctenopharyngodon idellus, NCCRP-1, Clone Prokaryotic expression

蔡佳, 代礼平, 王蓓, 汤菊芬, 黄郁葱, 鲁义善, 吴灶和, 简纪常. 草鱼(Ctenopharyngodon idellus)NCCRP-1基因的克隆和原核表达[J]. 生物技术通报, 2013, 0(11): 105-111.

Cai Jia, Dai Liping, Wang Bei, Tang Jufen, Huang Yucong, Lu Yishan, Wu Zaohe, Jian Jichang. cDNA Cloning and Prokaryotic Expression of NCCRP-1 from Grass Carp Ctenopharyngodon idellus)[J]. Biotechnology Bulletin, 2013, 0(11): 105-111.

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草鱼(Ctenopharyngodon idellus)NCCRP-1基因的克隆和原核表达

cDNA Cloning and Prokaryotic Expression of NCCRP-1 from Grass Carp Ctenopharyngodon idellus)

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[1]	吕秋谕, 孙培媛, 冉彬, 王佳蕊, 陈庆富, 李洪有. 苦荞转录因子基因FtbHLH3的克隆、亚细胞定位及表达分析[J]. 生物技术通报, 2023, 39(8): 194-203.
[2]	王佳蕊, 孙培媛, 柯瑾, 冉彬, 李洪有. 苦荞糖基转移酶基因FtUGT143的克隆及表达分析[J]. 生物技术通报, 2023, 39(8): 204-212.
[3]	孙明慧, 吴琼, 刘丹丹, 焦小雨, 王文杰. 茶树CsTMFs的克隆与表达分析[J]. 生物技术通报, 2023, 39(7): 151-159.
[4]	梅欢, 李玥, 刘可蒙, 刘吉华. 小檗碱桥酶高效原核表达及生物合成l-SLR的研究[J]. 生物技术通报, 2023, 39(7): 277-287.
[5]	郭三保, 宋美玲, 李灵心, 尧子钊, 桂明明, 黄胜和. 斑地锦查尔酮合酶基因及启动子的克隆与分析[J]. 生物技术通报, 2023, 39(4): 148-156.
[6]	杨俊钊, 张新蕊, 赵国柱, 郑菲. 新型GH5家族多结构域纤维素酶的结构与功能研究[J]. 生物技术通报, 2023, 39(4): 71-80.
[7]	刘思佳, 王浩楠, 付宇辰, 闫文欣, 胡增辉, 冷平生. ‘西伯利亚’百合LiCMK基因克隆及功能分析[J]. 生物技术通报, 2023, 39(3): 196-205.
[8]	陈楚雯, 李洁, 赵瑞鹏, 刘媛, 吴锦波, 李志雄. 藏鸡GPX3基因的克隆、组织表达谱研究及功能预测[J]. 生物技术通报, 2023, 39(3): 311-320.
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[10]	侯瑞泽, 鲍悦, 陈启亮, 毛桂玲, 韦博霖, 侯雷平, 李梅兰. 普通白菜PRR5的克隆、表达及功能验证[J]. 生物技术通报, 2023, 39(10): 128-135.
[11]	杨敏, 龙雨青, 曾娟, 曾梅, 周新茹, 王玲, 付学森, 周日宝, 刘湘丹. 灰毡毛忍冬UGTPg17、UGTPg36基因克隆及功能研究[J]. 生物技术通报, 2023, 39(10): 256-267.
[12]	郭文博, 路杨, 隋丽, 赵宇, 邹晓威, 张正坤, 李启云. 球孢白僵菌真菌病毒BbPmV-4外壳蛋白多克隆抗体制备及应用[J]. 生物技术通报, 2023, 39(10): 58-67.
[13]	陈光, 李佳, 杜瑞英, 王旭. 水稻盐敏感突变体ss2的鉴定与基因功能分析[J]. 生物技术通报, 2022, 38(9): 158-166.
[14]	孙威, 张艳, 王聿晗, 徐僡, 徐小蓉, 鞠志刚. 马缨杜鹃Rd3GT1的克隆及对矮牵牛花色形成的影响[J]. 生物技术通报, 2022, 38(9): 198-206.
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