摘要
根据已知罗氏沼虾(Macrobrachium rosenbergii)雌激素相关受体基因序列(Gen Bank登录号KU899089),设计含有酶切位点的特异性引物,PCR扩增得1 374 bp的开放阅读框序列,构建原核表达载体p ET-32a-ERR,并优化诱导浓度、诱导时间、诱导温度等表达条件。结果表明,ERR蛋白在温度35℃、IPTG浓度0.1 mmol/L条件下,诱导4 h时表达量较佳。
Estrogen-related receptor (ERR) belongs to the orphan nuclear receptor family, which mainly participates in the estrogen signaling pathway, and plays key role in the regulation of metabolism and development. According to the ERR sequences of Macrobrachium rosenbergii (GenBank accession number: KU899089), the specific primers containing restriction sites was designed and synthesized. The open reading frame (ORF) of ERR was amplified by PCR, and was constructed into the pET-32a vector for the prokaryotic expression. Concentration of IPTG, time of induction and temperature were optimized for ERR protein expression. Results showed that the ERR protein could be successfully and efficiently expressed under the condition of 0.1 mmol/L IPTG, and 35 ℃ for 4 hours in Escherichia coli.
作者
赵苗鑫
陈华谱
刘金磊
邓思平
李广丽
朱春华
洪宇聪
ZHAO Miao-xin;CHEN Hua-pu;LIU Jin-lei;DENG Si-ping;LI Guang-li;ZHU Chun-hua;HONG Yu-cong(Zhanjiang City State Key Laboratory of Marine Ecology and Environment, Fisheries College,Guangdong Ocean University, Zhanjiang 524088, China;Guangdong ZhenYuan Aquatic Industry Co.,Jieyang 522010, China)
出处
《广东海洋大学学报》
CAS
2017年第1期108-112,共5页
Journal of Guangdong Ocean University
基金
广东省海洋渔业科学与技术推广项目
(B201500B02)
广东省科技计划项目(2014B090905008)
广东海洋大学科学项目(E15181)
