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Short Hydrogen Bonds and Proton Delocalization in Green Fluorescent Protein (GFP)
Short Hydrogen Bonds Proton Delocalization Green Fluorescent Protein GFP
2016/5/25
Short hydrogen bonds and specifically low-barrier hydrogen bonds (LBHBs) have been the focus of much attention and controversy for their possible role in enzymatic catalysis. The green fluorescent pro...
Ground-state Proton Transfer Kinetics in Green Fluorescent Protein (GFP)
Mitotic centromere-associated kinase (MCAK/Kif2C) Cellular senescence p53 Human primary cells
2016/5/25
Proton transfer plays an important role in the optical properties of green fluorescent protein (GFP). While much is known about excited-state proton transfer reactions (ESPT) in GFP occurring on ultra...
GFP Variants with Alternative β‑Strands and Their Application as Light-driven Protease Sensors:A Tale of Two Tails
Green Fluorescent Proteins Peptide Hydrolases Light
2016/5/25
Green fluorescent protein (GFP) variants that carry one extra strand 10 (s10) were created and characterized, and their possible applications were explored. These proteins can fold with either one or ...
Thermodynamics,Kinetics,and Photochemistry of Beta-Strand Association and Dissociation in a Split-GFP System
Green Fluorescent Proteins Protein Structure,Secondary Kinetics Photochemistry Thermodynamics Models, Molecular
2016/5/25
Truncated green fluorescent protein (GFP) that is refolded after removing the 10th β-strand can readily bind to a synthetic strand to recover the absorbance and fluorescence of the whole protein. This...
The Electronic Structure of the Chromophore in Green Fluorescent Protein (GFP)
Electronic Structure Chromophore Green Fluorescent Protein GFP
2016/5/23
The chromophore responsible for the fluorescence from Green Fluorescent Protein (GFP) of the jelly fish Aequorea Victoria1,2 is formed in an autocatalytic, posttranslational cyclization and oxidation ...
Development and Characterization of Caspase Activatable GFP and a Family of Fluorescent Reporters
apoptosis GFP
2014/11/20
The cellular process of programmed cell death, or apoptosis, is critical in homeostasis and development. In addition it's misfunction is implicated in an array of disease states from cancer to neurode...
为了研究不同表达系统获得的携带绿色荧光抗HER2单链抗体(Anti-HER2-ScFv-GFP)是否既可靶向结合HER2阳性乳腺癌细胞表面, 也可通过观察绿色荧光变化直接判断抗体结合乳腺癌细胞表面后细胞的动态变化, 在前期成功构建两种表达系统的基础上, 利用Ni2+-NTA亲和层析法纯化来源于真核表达系统pFAST Bac to Bac HT A/Tn-5B1-4和原核表达系统pBAD His B...
通过对大麦小孢子进行基因枪轰击获得4 株转绿色荧光蛋白基因(gfp)的植株(A、C、D、E ),以gfp基因为探针进行荧光原位杂交( FISH)研究转化植株中转基因插入位置和基因表达。4 个株系在染色体7L(5HL)的不同位置都有一个插入点,而E 株系在染色体5S ( 7HS)还有第2个插入点。所有的转基因T0代植株都是半合子并在T1、T2代发生分离。D 株系GFP未表达,但FISH 和PCR分析...
利用GFP示踪细胞内源性P53活性检测DNA损伤
绿色荧光蛋白 P53 DNA损伤 检测
2008/1/7
DNA损伤的检测对预防癌症和遗传病等非常重要。采用分子克隆技术,将报告基因―绿色荧光蛋白(GFP)置于SV40基本启动子调控下,构建成对照载体pSV-GFP。在SV40基本启动子上游插入寡核苷酸P53RE,构建成示踪载体p53RE-GFP。转染NIH 3T3细胞,以GFP示踪细胞内源性P53的转录激活活性。紫外线照射或H2O2处理转化细胞使DNA损伤,诱导细胞内源性P53的表达。用激光扫描共聚焦成...
以绿色荧光蛋白GFP基因为报道基因,用花粉管通道和农杆菌介导的转化方法将外源基因导入棉花(Gossypium hirsutumL.),分别获得转化幼胚、幼苗和转化愈伤组织。用手持紫外灯结合显微镜检术能够快速地对转化子进行活体筛选鉴定,比用GUS检测方法有明显的优越性。本研究不但为花粉管通道转化法的可行性提供了新的证据,同时也建立了GFP用于棉花基因工程研究的检测技术体系。
用绿色荧光蛋白(GFP)作为报告分子筛选有效的siRNA
RNA干扰 荧光定量PCR 绿色荧光蛋白 载体
2008/1/3
摘要 摘要 建立一种利用绿色荧光蛋白(GFP)作为报告分子筛选能有效抑制目的基因表达的siRNA的方法.以巨噬细胞移动抑制因子(MIF)基因为研究对象,筛选能有效沉默MIF表达的质粒载体介导的siRNA.构建拥有同一Kozak共有翻译启始序列、翻译启始密码子ATG的MIF-GFP融合表达载体pEGFP-MIF.分别将3个靶向MIF的siRNA表达质粒与pEGFP-MIF共转化HEK293细胞,在...
GFP标记在转基因青鳉同系繁殖纯化中的应用Application of GFP Mark on Inbred Depuration of Transgenic Medaka (Oryzias latipes)
绿色荧光蛋白(GFP)基因 显微注射 青鳉 同系繁殖 纯化
2007/12/31
摘要从载体pBluescript SK+的Nde I位点处插入外源基因片段,构建了含有青鳉(Oryzias latipes)β-肌动蛋白(β-Actin)启动子和绿色荧光蛋白(GFP)标记基因的新表达载体。显微注射实验证实:在青鳉受精卵的单细胞期进行显微注射,标记基因的表达率和胚胎的存活率均较高。对嵌合体的转基因青鳉(F1)进行了两代的杂合体(F2和F3)筛选和一代的纯合体(F4)筛选,获得第4代...
摘要 为研究内生细菌对宿主植物侵染定殖的机理和其共生生物学作用 ,对水稻内生优势成团泛菌 (Pantoeaagglomerans)YS19与绿色荧光蛋白 (GFP)标记的YS19B ::gfp菌株的生长动力学进行了比较研究 ,探讨了成团泛菌YS19B ::gfp的标记稳定性和荧光性质 .标记菌株与野生型菌株相比 ,最大比生长速率和最大生物量仅减小 12 4 %和 6 % ,代时延长 14 0 %...
摘要
为了用绿色荧光蛋白标记观察人类无精症相关基因ZNF230在Cos7细胞中的蛋白质表达及定位,用PCR方法扩增得到突变的人和小鼠mt-ZNF230和mt-znf230基因,使其3′端的终止密码TGA突变为TGG,并装入T-载体,双酶切后通过定向克隆将其与真核表达载体pEGFP-N1的绿色荧光蛋白(green fluorescence protein,GFP)基因融合,构建了ZNF230—荧光...
GFP为报告基因的酵母哺乳动物细胞穿梭载体的构建及其在人血管内皮细胞中的表达
酵母 穿梭载体 口服基因治疗
2007/12/19
摘要 为研究酵母作为载体在口服基因治疗及免疫中的作用 ,需要一种能够在酵母中复制而在哺乳动物细胞中表达的穿梭载体 .利用通用载体质粒融合系统 (UPS)构建了一种以GFP为报告基因的新载体 ,以常规的氯化锂法对酿酒酵母进行转化 ,证明该载体能够在酵母中复制 ;以脂质体介导向人血管内皮细胞进行了转染 ,有绿色荧光 ,证明该载体能够在哺乳动物细胞中表达 .所获得的新型的穿梭载体为口服酵母在基因治疗中的...