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荧光共振能量转移(FRET)体系的构建及其验证 被引量:1
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作者 钱月娇 冯钰斌 +4 位作者 钱学文 朱传君 李鸽 卢多 陈飞虎 《安徽医科大学学报》 CAS 北大核心 2018年第12期1821-1827,共7页
目的构建携带FRET体系(YFP-CFP对)的一组原核表达工程质粒,在大肠杆菌(E. coli)中高效表达后,验证该体系FRET信号作为细胞内监测蛋白相互作用指标的可行性。方法分别构建N端、C端以及两端分别编码YFP和(或) CFP的6个原核表达工程质粒:pN... 目的构建携带FRET体系(YFP-CFP对)的一组原核表达工程质粒,在大肠杆菌(E. coli)中高效表达后,验证该体系FRET信号作为细胞内监测蛋白相互作用指标的可行性。方法分别构建N端、C端以及两端分别编码YFP和(或) CFP的6个原核表达工程质粒:pNYFP、pCYFP、pNCFP、pCCFP、pYFP-CFP、pCFP-YFP。设计YFP-CFP和CFP-YFP作为阳性对照,而等量的YFP与CFP混合的体系(YFP+CFP)作为阴性对照。将过表达的工程蛋白通过镍柱、分子筛层析柱进行初步纯化后使用酶标仪检测相同摩尔浓度纯化蛋白的FRET信号。在此基础上,进一步用酶标仪检测表达工程蛋白的菌液中的FRET信号。结果通过测序验证,成功构建了6个原核表达的工程质粒。酶标仪检测的结果显示在体外纯化蛋白条件和菌液条件下均可产生明确的FRET信号。结论成功构建了FRET体系的原核表达工程质粒,不仅验证了蛋白条件下的可行性,而且验证了菌液条件下的可行性,为在活细胞条件下对蛋白-蛋白相互作用进行实时的动态研究,提供一个非常便利的实验方法。 展开更多
关键词 荧光共振能量转移 蛋白-蛋白相互作用 活细胞 原核表达 工程质粒 蛋白纯化
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An analytical model for pyrolysis of a single biomass particle 被引量:4
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作者 Mehdi Bidabadi Mohammad Rastegar Moghaddam +2 位作者 Seyed Alireza Mostafavi Farzad Faraji Dizaji Hossein Beidaghy Dizaji 《Journal of Central South University》 SCIE EI CAS CSCD 2015年第1期350-359,共10页
Decreasing in emissions of greenhouse gases to confront the global warming needs to replace fossil fuels as the main doer of the world climate changes by renewable and clean fuels produced from biomass like wood waste... Decreasing in emissions of greenhouse gases to confront the global warming needs to replace fossil fuels as the main doer of the world climate changes by renewable and clean fuels produced from biomass like wood waste which is neutral on the amount of CO2. An analytical and engineering model for pyrolysis process of a single biomass particle has been presented. Using a two-stage semi global kinetic model which includes both primary and secondary reactions, the effects of parameters like shape and size of particle as well as porosity on the particle temperature profile and product yields have been investigated. Comparison of the obtained results with experimental data shows that our results are in a reasonable agreement with previous researchers' works. Finally, a sensitivity analysis is done to determine the importance of each parameter on pyrolysis of a single biomass particle which is affected by many constant parameters. 展开更多
关键词 biomass pyrolysis analytical model single particle sensitivity analysis
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