X

Download The Yeast Two Hybrid System The Jena Protein Protein PowerPoint Presentation

SlidesFinder-Advertising-Design.jpg

Login   OR  Register
X


Iframe embed code :



Presentation url :

Home / Health & Wellness / Health & Wellness Presentations / The Yeast Two Hybrid System The Jena Protein Protein PowerPoint Presentation

The Yeast Two Hybrid System The Jena Protein Protein PowerPoint Presentation

Ppt Presentation Embed Code   Zoom Ppt Presentation

PowerPoint is the world's most popular presentation software which can let you create professional The Yeast Two Hybrid System The Jena Protein Protein powerpoint presentation easily and in no time. This helps you give your presentation on The Yeast Two Hybrid System The Jena Protein Protein in a conference, a school lecture, a business proposal, in a webinar and business and professional representations.

The uploader spent his/her valuable time to create this The Yeast Two Hybrid System The Jena Protein Protein powerpoint presentation slides, to share his/her useful content with the world. This ppt presentation uploaded by worldwideweb in Health & Wellness ppt presentation category is available for free download,and can be used according to your industries like finance, marketing, education, health and many more.

About This Presentation

Slide 1 - The Yeast Two-Hybrid System Anne C. Luebke
Slide 2 - What is the yeast two-hybrid system used for? Identifies novel protein-protein interactions Can identify protein cascades Identifies mutations that affect protein-protein binding Can identify interfering proteins in known interactions (Reverse Two-Hybrid System)
Slide 3 - How does it work? Uses yeast as a model for eukaryotic protein interactions A library is screened or a protein is characterized using a bait construct Interactions are identified by the transcription of reporter genes Positives are selected using differential media
Slide 4 - The Model DNA-Binding Domain Bait Protein Prey Protein Transcription Activating Region Reporter Gene DNA-Binding Site
Slide 5 - Steps to Screen a Library Create the Bait Plasmid Construct from the gene of interest and the DNA binding domain of Gal4 or LexA or other suitable domain Transform with the bait construct a yeast strain lacking the promoter for the reporter genes and select for transformed yeast Transform the yeast again with the library plasmids Select for interaction
Slide 6 - Sequence analysis Isolate plasmid from yeast and transform E. coli Purify plasmid from E. coli and sequence Blast sequence against database for known proteins or construct a possible protein sequence from the DNA sequence and compare to other proteins
Slide 7 - Reporter Genes LacZ reporter - Blue/White Screening HIS3 reporter - Screen on His+ media (usually need to add 3AT to increase selectivity) LEU2 reporter - Screen on Leu+ media ADE2 reporter - Screen on Ade+ media URA3 reporter - Screen on Ura+ media (can do negative selection by adding FOA)
Slide 8 - Plasmid Constructs Plasmids are constructed with the Gal4 DNA binding domain (or other suitable domain) in front of a Multiple Cloning Site (MCS) The plasmid contains genes that can be used for selection such as Amp, Leu2, Ura3, or Trp1
Slide 9 - Sample Plasmid From Golemis Lab Homepage
Slide 10 - False Positives False positives are the largest problem with the yeast two-hybrid system Can be caused by: Non-specific binding of the prey Ability to induce transcription without interaction with the bait (Majority of false positives)
Slide 11 - Elimination of False Positives Sequence Analysis Plasmid Loss Assays Retransformation of both strain with bait plasmid and strain without bait plasmid Test for interaction with an unrelated protein as bait Two (or more) step selections
Slide 12 - Advantages Immediate availability of the cloned gene of the interacting protein Only a single plasmid construction is required Interactions are detected in vivo Weak, transient interactions can be detected Can accumulate a weak signal over time
Slide 13 - Examples of Uses of the Yeast Two-Hybrid System Identification of caspase substrates Interaction of Calmodulin and L-Isoaspartyl Methyltransferase Genetic characterization of mutations in E2F1 Peptide hormone-receptor interactions Pha-4 interactions in C. elegans
Slide 14 - References Bartel, Paul, C. Chien, R. Sternglanz, S. Fields. “Elimination of False Positives that Arise in Using the Two-Hybrid System.” Biotechniques (1993) Vol. 14, no. 6, p. 920-924. Chien, Cheng-ting, P. Bartel, R. Sternglanz, S. Fields. “The two-hybrid system: A method to identify and clone genes for proteins that interact with a protein of interest.” Proc. Natl. Acad. Sci. USA (1991) Vol. 88, p. 9578-9582. Fields, Stanley, O. Song. "A novel genetic system to detect protein-protein interactions." Nature (1989) Vol. 340, p.245-246. James, Philip, J. Halladay, E. Craig. "Genomic Libraries and a Host Strain Designed for Highly Efficient Two-Hybrid Selection in Yeast." Genetics (1996) Vol. 144, p. 1425-1436. Kamada, S, H. Kusano, H. Fujita, M. Ohtsu, R. Koya, N. Kuzumaki, Y. Tsujimoto. "A cloning method for caspase substrates that uses the yeast two-hybrid system: Cloning of the antiapoptotic gene gelsolin." Proc. Natl. Acad. Sci. USA (1998) Vol 95, p. 8532-8537. O'Connor, Mirriam, C. O'Connor. "Complex Interactions of the Protein L-Isoaspartyl Methyltransferase and Calmodulin Revealed with the Yeast Two-hybrid System." The Journal of Biological Chemistry (1998) Vol. 273, p. 12909-12913. Staudinger, Jeff, J. Zhou, R. Burgess, S. Elledge, E. Olson. "PICK1: A Perinuclear Binding Protein and Substrate for Protein Kinase C Isolated by the Yeast Two-Hybrid System." The Journal of Cell Biology (1995) Vol. 128, p. 263-271.
Slide 15 - References continued Vidal, Marc, P. Braun, E. Chen, J. Boeke, E. Harlow. "Genetic Characterization of a mammalian protein-protein interaction domain by using a yeast reverse two-hybrid system." Proc. Natl. Acad. Sci. USA (1996) Vol. 93, p. 10321-10326. White, Michael. "The yeast two-hybrid system: Forward and reverse." Proc. Natl. Acad. Sci. USA (1996) Vol 93, p. 10001-10003. Zhu, Jianwei, C. Kahn. "Analysis of a peptide hormone-receptor interaction in the yeast two-hybrid system." Proc. Natl. Acad. Sci. USA (1997) Vol. 94, p. 13063-13068. Lab of Erica Golemis http://www.fccc.edu/research/labs/golemis/EG_homepage.html Special thanks to Dr. Susan Mango and the University of Utah
 

Frequently Asked Questions

> About SlidesFinder?

Slidesfinder is a website that allows users to upload and share their PowerPoint presentations online. It provides a platform for individuals and businesses to share their ideas and knowledge through presentations, and for others to view and download these presentations for educational or professional purposes.
Slidesfinder offers a wide range of presentation topics and categories, including business, education, science, technology, health, and more. Users can also search for presentations using keywords, tags, or by category. The website provides tools to create professional presentations with rich media, animations, and graphics.



One of the key features of Slidesfinder is its user-friendly interface, which allows users to easily upload, view, and share presentations. Users can also embed their presentations in their websites or blogs using the website's embed code feature.
Slidesfinder also offers premium services, such as the ability to create private presentations and to download presentations in high-quality formats. However, the basic features of Slidesfinder are free to use, making it accessible to anyone who wants to share their ideas and knowledge through presentations.

> How do I register with SlidesFinder?

Go to registration page (you can see signup link on top of website page) https://www.slidesfinder.com/signup . If you have facebook/gmail account them just click on SIGN IN WITH FACEBOOK OR SIGN IN WITH GOOGLE button, by this you will be a registered member of slidesfinder without filling any form, required detail automatically will be fatch from your account. If you do not a Facebook account, then click on "Signup". Fill all required fields and you will be a registered member of slidesfinder.

> Is slidesfinder account confirmation is mandatory?

Yes it is mandetory to active your account to login.

> Do I need to signup/login on SlidesFinder before uploading a PowerPoint presentation?

Yes, you need to login with your account before uploading presentation. Your username will be displayed on your uploaded presentation. Your registered email id is needed for sending your stats of uploaded presentation.

Slidesfinder is a sharing website for PowerPoint presentations search and share. Find your interest in the form of powerpoint presentations on slidesfinder and save your valuable time . On Slidesfinder you get presentations from our huge library of professional ppt presentations. We believe in making your search INFORMATIVE and FUN. Find your best ppt presentation from a pool of PowerPoint presentations stacked under important industry categories like business & management, heath & Wellness,eduction & training etc. We provide unique informative PowerPoint presentation for marketers, presenters and educationists. These professional PowerPoint presentations are uploaded by professionals from across numerous industry segments.These ppt presentations are available for FREE download.

Not just finding your interest, but facilitate you broadcast your interest. We have created this platform for easy sharing of PowerPoint presentations, ensuring that these presentations get maximum exposure. Create your slidesfinder account and upload PowerPoint presentations for free, share on social media platforms and BUILD YOUR CROWD WITH PRESENTATION !!

The Great Buddha says, "Share your knowledge.It’s a way to achieve immortality"! So, start sharing knowledge and we are here to make that immortal !!

© 2013-2024 SlidesFinder. All rights reserved.

Re-productivity of content are not allowed. Without prior written permission from author, commercial use of any content is illegal.