云南农业大学学报（自然科学）2017，Vol.32Issue(2)：191-196,6.DOI:10.16211/j.issn.1004-390X(n).2017.02.001
纳米荧光素酶的表达、纯化及性质鉴定
Expression, Purification and Characterization of Nano Luciferase in Escherichia coli
摘要
Abstract
A large group of luminescence reactions catalyzed by luciferase are called bioluminescence.Since bioluminescence has been characterized as low background,high sensitivity and wide linear dynamic range.It is applied widely in sensitive molecular biology detection.In this research,we have synthesized a novel luciferase (nano luciferase) by whole gene synthesis,to make it a novel bioluminescent detection tool.Nano luciferase was cloned into a prokaryotic expression vector and expressed in an auto-induced system.Recombinant nano luciferase was purified by Ni-NTA affinity chromatography and size exclusive chromatography.The typical yield of recombinant nano luciferase was 25.3 mg from a liter culture medium with purity above 98％.Then,we tested the activity of recombinant nano luciferase,from 1.0 × 10-4 mg/mL to 1.0 × 10-7 mg/mL,nano luciferase has good linear response,the detection limit was as low as 1.0 × 10-7 mg/mL (around 5 pmol).The assay based on nano luciferase is ultrasensitive,good repeatable and easy operated.It would be applied as a basic tool to develop new sensitive bioluminescent probes with promising novel applications.关键词
纳米荧光蛋白/原核表达/纯化Key words
nano luciferase (NLuc)/prokaryotic expression/purification分类
生物科学引用本文复制引用
侯家鹏,丁澦..纳米荧光素酶的表达、纯化及性质鉴定[J].云南农业大学学报（自然科学）,2017,32(2):191-196,6.基金项目
国家自然科学基金项目(31470764). （31470764）
