Tel:4001790116,13012892256,17521589219

Cell, culture reagent, life science research overall service provider!

Technical Support技术支持

CONTACT US

Application And Carrier System Of GFP Marker

source:Qida organism  views:2271  time:2021-06-18

The Bioluminescence And Fluorescence Of Green Fluorescent Protein (GFP) And Other Proteins May Have Existed In Jellyfish And Other Organisms For Millions Of Years; However, It Was Not Until The 1960s That Scientists Began To Study GFP And Infer Its Biochemical Characteristics. Now, GFP And Its Fluorescent Derivatives Have Become The Main Research Objects In The Laboratory. GFP Is Widely Used In Biological Research. Scientists Use GFP To Achieve A Variety Of Functions, Including: Labeling Genes To Clarify Their Expression Or Location Characteristics, Acting As Biosensors Or Cell Markers, Studying The Interaction Between Proteins, Visualizing The Activity Of Promoters, And So On. Why Green Fluorescent Protein& Nbsp; GFP Is A Protein Of About 27 Kda, Which Is Composed Of 238 Amino Acids And Is Derived From The Crystal Jellyfish Aequorea Victoria. It Has Fluorescence Emission Wavelength (hence Its Name) In The Green Part Of The Visible Spectrum, Which Is Due To The Mature Reaction Of Three Specific Amino Acids (Ser65, Tyr66 And Gly67) In The Center Of The Protein. When It Was First Discovered, One Of The Most Surprising Aspects Of GFP Was That Chromophores Were Formed Spontaneously, Without Additional Cofactors, Substrates Or Enzyme Activities, And It Only Needed To Exist In Oxygen During Maturation. This Means That Proteins Can Be Directly Expressed From A To B In Any Organism While Still Maintaining Fluorescence& Nbsp; The Protein Structure First Reported In 1996 Was 11 β- Contains "Barrel" Shaped Sheets, With Chromophores Hidden In The Center Of The Structure And Not Quenched By Aqueous Solution. This Tightly Arranged Structure Explains The Importance Of The Entire GFP Protein, Which Is Almost Completely Necessary To Maintain Fluorescence Activity; Compared With The Traditional Fluorescent Dyes At That Time, The Main Advantage Of GFP Is That It Is Non-toxic And Can Be Expressed In Living Cells, So That Dynamic Physiological Processes Can Be Studied& Nbsp; Redesign GFP To Increase Its Color And Application Range Almost After Its Sequence Was Clarified, Scientists Began To Modify The New Version Of GFP Through Mutation To Improve Its Physical And Biochemical Properties. In 1995, Roger Y. Tsien Describes A S65T Point Mutation That Increases The Fluorescence Intensity And Photostability Of GFP. This Also Shifted Its Main Excitation Light From 395 Nm To 488 Nm, Effectively Improving The Defects Of Wild Type Protein, And Promoting Its Wide Application In Research. Many Other Mutations Have Been Introduced Into GFP, And New Fluorophores Have Been Designed. Table 1 Below Lists Several Common Fluorescent Proteins And Their Mutations Relative To Wild-type GFP. Although Not Listed Here, Many Penetrations Exist In Each Color, With Only Slight Differences Between Them& Nbsp; Please Note That Many Fluorescent Proteins Found In The Red Part Of The Spectrum Are Not GFP Derivatives, But Related To The DsRed Protein Isolated From Discosoma Sp. Similar Work Has Been Done To Expand The Red Fluorescent Protein Library; However, These Proteins Are Unique Green Fluorescent Proteins And The Mutation Definition Found In Table 2 May Not Be Applicable. Table 1: Specific Mutations Containing Common Fluorophores; Table 2: Function Of Specific Mutations Of GFP Derivatives; Due To GFP And Its Ease Of Use, GFP And Other Fluorescent Proteins Have Become The Mainstream Of Molecular Biology. Scientists Can Easily Use GFP Containing Plasmids As A Means To Achieve Many Functional Purposes. The Following Are Some Commonly Used Uses, But There Are Many Other Uses At Present, And New GFP Technology Is Constantly Developing& Nbsp; Fusion Marker: GFP Is One Of The Most Common Uses. It Can Fuse With The N - Or C-terminal Of Protein, Which Enables Scientists To Visualize The Time And Place Of Gene Expression& Nbsp; Transcription Reporter: Placing GFP Under The Control Of The Promoter Of Interest Can Be Used To Effectively Monitor The Gene Expression Of The Promoter In Specific Cell Types. This Transcriptional Report Is One Of The Earliest Applications Of GFP& Nbsp; F ö Rster Resonance Energy Transfer (FRET): This Is Used To Study The Interaction Between Two Proteins Or The Interaction Between Two Protein Domains With Conformational Changes. Typically, Two Fluorescent Proteins With Overlapping Excitation/emission Spectra Are Used; One Fused To Each Protein Or Domain Being Tested. Here We Find The Micromotion Plasmid& Nbsp; Split EGFP: As An Alternative To Fet, Split EGFP Is Also Used To Study Protein Protein Interactions. In This Case, The Two Parts Of EGFP Fuse Into The Protein Of Interest. When They Are Close, The Two Parts Of EGFP Undergo Folding, Maturation And Fluorescence& Nbsp; Biosensors: A Series Of GFP Based Fluorescent Biosensors Have Been Designed Using Multiple Strategies Such As FRET And Calmodulin To Detect Various Intracellular Conditions, Including Ion (such As Ca2+) Concentration And PH, Etc. Photogenetics: Scientists Can Use Light To Detect, Measure And Control Molecular Signals, Cells And Cell Populations To Understand Their Activities And Visualize The Impact Of Changes On Such Activities. Learn More About Open Optogenetics In Optogenetics, And Find Photoactuators And Sensors In Addgene& Nbsp; Cell Marker/selection: Expression Structures Such As Plasmids Usually Include GFP As A Marker To Help Identify Which Cells Have Successfully Obtained Plasmids. This Can Be Used As An Alternative To Choosing Antibiotics. This Type Of Plasmid May Express GFP Under The Control Of The Additional Promoter Of The Gene Of Interest, Or The Same Transcript Of The Gene Of Interest, But After The Internal Ribosome Entry Site (IRES). This Is Typically Used For Connection To FACS (see Below)& Nbsp; Fluorescent Activated Cell Sorting (FACS): This Is A Flow Cytometry, Which Separates The Cell Mixture Into Different Groups According To The Fluorescence Signal. Therefore, FACS Can Be Used To Separate GFP Expressing Cells From Non GFP Expressing Cells& Nbsp; Developmental/transgenic Use: GFP Can Be Used For Genetic Tracking In Cell Fate Research Due To Its Stability. When Interest Promoters Control, It Can Also Be Used To Visually Show The Active Development Stage Of These Promoters. In Addition, GFP Can Also Label Transgenic ES Cells, Which Can Then Be Used For The Implantation And Generation Of Transgenic Mice& Nbsp; Purification: GFP Can Be Used As A General Epitope Marker For Protein Purification, And A Large Number Of Commercial Antibodies Against GFP Can Be Obtained& Nbsp; Other: We Just Scratched The Surface Of The Potential Application Of GFP. It Is Also Used To Identify Specific Cell Populations In Drug Screening, Visualize Micrometastasis In Nude Mice In Cancer Research, Act As A Reporter For DNA Double Strand Break Repair, And Mark Pathogenic Intracellular Microorganisms To Visualize Host/pathogen Interactions.
×