军事医学2012，Vol.36Issue(4)：253-257,262,6.
环式增强型绿色荧光蛋白热稳定性及Npu DnaE内含肽高反式剪接效率研究
Thermostability of cyclic EGFP and highly efficient protein trans-splicing of Npu DnaE intein
叶丙雨 1王政 2李德彬 2张乐之 2肖丽霞 2张部昌 2曹诚 1赵志虎2
作者信息
- 1. 安徽大学生命科学学院,合肥,230601
- 2. 军事医学科学院生物工程研究所,北京,100071
- 折叠
摘要
Abstract
Objective To produce Cyclic enhanced green fluorescent protein ( Cyc-EGFP) sre vivo using the highly efficient protein trans-splicing of Npu DnaE intein. Methods The fusion expression vector pET/Npu DnaE intein/EGFP-28a (+) was induced and expressed , and then using affinity purification high purity Cyc-EGFP and EGFP obtained , finally the fluorescence intensity and thermostability were compared. Results The trans-splicing activity of Npu DnaE intein had an efficiency above 98% , which was superior to that of Ssp DnaE intein. The Cyc-EGFP was 4 - 5^ more thermostable than the linear form. Conclusion The obvious advantage of Cyc -EGFP makes protein cyclization highly applicable to protein en - gineering and other fields.关键词
蛋白环化/内含肽类/Cyc-EGFP/反式剪接/热稳定性Key words
protein cyclization/ inteins/ Cyc-EGFP/ trans-splicing/ thermostability分类
生物科学引用本文复制引用
叶丙雨,王政,李德彬,张乐之,肖丽霞,张部昌,曹诚,赵志虎..环式增强型绿色荧光蛋白热稳定性及Npu DnaE内含肽高反式剪接效率研究[J].军事医学,2012,36(4):253-257,262,6.
