文献类型：期刊文献

中文题名：尼罗罗非鱼Galectin-4基因的原核表达及诱导条件优化

英文题名：Prokaryotic Expression and Optimization of Galectin4 Gene from Nile Tilapia(Oreochromis niloticus)

作者：张志强[1];刘鑫潮[1];牛金中[1];黄瑜[1];王蓓[1];简纪常[1]

机构：[1]广东海洋大学水产学院//广东省水产经济动物病原生物学及流行病学重点实验室//广东省水产经济动物病害控制重点实验室,广东湛江524088

年份：2020

卷号：40

期号：5

起止页码：118

中文期刊名：广东海洋大学学报

外文期刊名：Journal of Guangdong Ocean University

收录：CSTPCD

基金：国家自然科学基金青年基金项目(31702386);广东省科技厅国际科技合作领域项目(2017A050501037);广东省普通高校青年创新人才类项目(2018KQNCX105)。

语种：中文

中文关键词：尼罗罗非鱼;Galectin-4;基因克隆;原核表达;条件优化

外文关键词：Oreochromis niloticus;Galectin-4;gene clone;prokaryotic expression;condition optimization

中文摘要：【目的】研究尼罗罗非鱼(Oreochromisniloticus)Galectin-4基因(记作OnGal-4)的原核表达,为大量获取尼罗罗非鱼Galectin-4蛋白(OnGal-4)和后续功能研究奠定基础。【方法】根据NCBI上公布的尼罗罗非鱼Galectin-4基因序列(Genbank:XM-019345120)设计引物,构建原核表达载体,诱导Galectin-4重组蛋白(r OnGal-4)在BL21原核表达系统中表达,并优化其诱导条件。【结果与结论】成功扩增出OnGal-4基因,并构建原核表达载体pGEX-4T-On Gal-4。rOnGal-4最佳诱导条件是温度28℃,IPTG浓度0.4 mmol/L,诱导时间4 h。在此条件下,rOnGal-4在可溶性蛋白中大量表达。Western-blot结果显示,rOnGal-4可与抗GST标签的单克隆抗体发生特异性反应。

外文摘要：【Objective】To study the optimization conditions for prokaryotic expression of the Nile tilapia(Oreochromis niloticus)OnGal-4 gene,and perform functional studies of Galectin-4【.Method】A pair of primers were designed based on the tilapia Galectin-4 gene sequence(Genbank:XM-019345120),and to expression plasmid(pGEX-4T-OnGal-4)was produced.The Galectin-4 recombinant clone was induced to express rOnCal4 in BL21 cells and the expression conditions were optimized.【Result and Conclusion】The OnGal-4 gene was successfully amplified and the prokaryotic expression plasmid pGEX-4T-OnGal-4 was constructed.The optimal induction conditions for rOnGal-4 production were 0.4 mmol/L of IPTG at 28℃for 4 h.The rOnGal-4 protein was highly expressed under the optimized expression conditions.Western-blot results showed that the rOnGal-4 could specifically react with GST-tag monoclonal antibody.