首页> 外文期刊>Analytical methods >A novel molecular biosensor system model designed by using a couple composed of a fluorescent mutant binding protein and fluorophore-labeled ligand analogue based on the FRET mechanism
【24h】

A novel molecular biosensor system model designed by using a couple composed of a fluorescent mutant binding protein and fluorophore-labeled ligand analogue based on the FRET mechanism

机译:基于FRET机制,使用由荧光突变结合蛋白和荧光团标记的配体类似物组成的偶对设计的新型分子生物传感器系统模型

获取原文
           

摘要

Molecular biosensors represent a powerful form of biotechnology. In this study, a novel molecular biosensor model was designed based on fluorescence resonance energy transfer (FRET) that was carried out by using a couple composed of a genetically engineered fluorescent mutant binding protein and fluorophore-labeled ligand analogue. The pair of streptavidin and biotin was used as a model here. Trp120BFLAF mutant streptavidin (a fluorescent unnatural amino acid named BODIPY-FL-aminophenylalanine (BFLAF)) was position-specifically incorporated into the Trp120 position of streptavidin by a four-base codon method. The FRET occurred when the Alexa546-biotin was added into the Trp120 BFLAF mutant streptavidin. Furthermore, the FRET between Trp120BFLAF mutant streptavidin and Alexa546-biotin decreased by the competitive addition of natural biotin. This result demonstrated that by measuring the FRET, a couple composed of a genetically engineered fluorescent mutant binding protein (Trp120BFLAF mutant streptavidin) and fluorophore-labeled ligand analogue (Alexa546-biotin) was applicable for the quantification of the small ligand (free biotin) as a molecular biosensor system.
机译:分子生物传感器代表了一种强大的生物技术形式。在这项研究中,基于荧光共振能量转移(FRET)设计了一种新型分子生物传感器模型,该模型是通过使用由基因工程改造的荧光突变体结合蛋白和荧光团标记的配体类似物组成的一对进行的。这里以链霉亲和素和生物素对为模型。通过四碱基密码子方法将Trp120BFLAF突变型链霉亲和素(一种称为BODIPY-FL-氨基苯丙氨酸(BFLAF)的荧光非天然氨基酸)位置特异性地掺入到链霉亲和素的Trp120位。当将Alexa546-生物素添加到Trp120 BFLAF突变链霉亲和素中时,发生FRET。此外,通过竞争性添加天然生物素,Trp120BFLAF突变型链霉亲和素与Alexa546-生物素之间的FRET降低。该结果表明,通过测量FRET,由基因工程荧光突变结合蛋白(Trp120BFLAF突变链霉亲和素)和荧光团标记的配体类似物(Alexa546-生物素)组成的夫妇可用于定量小配体(游离生物素)。分子生物传感器系统。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号