ACADEMIA SINICA IBMS 嚴仲陽美國德州貝勒醫學院博士 Specialty:

Slides:

Advertisements

Similar presentations

Recombinant DNA Technology

Advertisements

Molecular biology tools for the genetic manipulation of yeast

BioBuilding: What a Colorful World. An engineering paradigm Design Build Test The focus of this lab.

Introduction of DNA into Living Cells

Biology Mathematics Engineering Optics Physics Robotics Informatics.

Yeast Two-Hybrid system for working with proteins affecting yeast growth Maarten de Smit

The Yeast Two-Hybrid System Anne C. Luebke. What is the yeast two-hybrid system used for? n Identifies novel protein-protein interactions n Can identify.

Molecular biology Transformation: introduction of DNA – Selectable marker – Spheroplasts, Li 2+ salts, electroporation Yeast plasmids are shuttle plasmids,

Preparation of competent E. coli cells Transformation of the ligated construct into E. coli Purification of the digested fragment Purification of the pBR322.

AP Biology Chapter 18: Gene Regulation. Regulation of Gene Expression Important for cellular control and differentiation. Understanding “expression” is.

Andres Alvarez Dr. Jeff Chang IDENTIFICATION OF CANDIDATE TARGET PROTEINS OF TYPE III EFFECTORS.

A comprehensive analysis of protein-protein interactions in Saccharomyces cerevisiae Article by Peter Uetz, et.al. Presented by Kerstin Obando.

Polymerase Chain Reaction - PCR The photocopier of molecular biology.

Expression of the lacZ Gene from the ibpB Heat Shock Protein Ebee Hornbeck & Sheena Donald.

Transposable Mutagenesis for an E. coli Methionine or Histidine Synthesis Gene Jon Kissel Mary Maschek.

DNA Science Day 2 Extracting, Ligating and Transforming Physical Biology Bootcamp October 2006 Caltech.

Sporulation and aquaporins in Saccharomyces cerevisiae

Protein-protein Interactions Hsueh-Fen Juan 2003, Mar 31 NTNU.

What I’ve done this summer Institute of Molecular Biology – Academia Sinica Dr. Che-Kun James Shen 沈哲鯤博士 National Health Research Institutes Dr. Xin Chen.

Genetics Laboratory BL415, Spring 2015 Yeast Transformation.

Protein-Protein Interaction Screens. Bacterial Two-Hybrid System selectable marker RNA polymerase DNA binding protein bait target sequence target.

研習報告與討論研習報告與討論蔡騰寬中研院生醫所林陽生老師實驗室. Interaction of TCF4 with DP103 and FHL3.

Preparation of competent E. coli cells Preparation of probes by PCR Digestion of pBR322 and plasmid with restrionenzymes BamH1, EcoR1 and EcoRV Purification.

MCB 317 Genetics and Genomics MCB 317 Topic 10, part 5 A Story of Transcription.

In Vivo assay for RNA-protein interactions Dana M. Schneider Loren Williams lab.

Goal: To identify yeast gene products important for accurate chromosome transmission in mitosis. Importance: Errors during chromosome transmission in humans.

Biology of genetic engineering Host cell types. Host cell types 1. Prokaryotic hosts; such as E. coli 2. Eukaryotic hosts; range from microbes, such as.

Genetic Engineering. Tools of Molecular Biology DNA Extraction Cutting DNA Restriction Enzymes Recognize certain sequences of DNA and cut the hydrogen.

Protein protein interactions

Recombinant DNA Technology Bacterial Transformation & GFP.

Seeds are mutagenized in the lab, then screened for mutants in the ethylene signaling pathway, based on the “triple response” phenotype. The mutants that.

BUDDING TECHNOLOGIES AND BUDDING YEAST 2012 HHMI Summer Workshop for High School Science Teachers.

(D) Crosslinking Interacting proteins can be identified by crosslinking. A labeled crosslinker is added to protein X in vitro and the cell lysate is added.

Two-Hybrid Vector Systems Detection of Protein-Protein Interactions.

Transient Protein-Protein Interactions (TPPI)

Ethylene responses Developmental processes

Yeast as a Model System MBIOS 520/420 September 29, 2005.

Creating an RNAi feeding vector How does ligation into L4440 work?

Suzanne Vroman Katze Lab University of Washington Protein-Protein Interactions of P58 IPK.

Bi Sternberg Analyzing and Building Chromosomes.

Discovering Macromolecular Interactions. An experimental strategy for identifying new molecular actors in a process candidate approach general screen.

Coupling teaching and research Incorporating a research project into a required course.

Protein-protein interactions “The Interactome” Yeast two-hybrid analysis Yeast two-hybrid analysis Protein chips Protein chips Biochemical purification/Mass.

LOGO Isolation and characterization of regulators of oxidative stress induced apoptosis in yeast Yaron Fireizen, Christine Crozier and Julie Anderson Biology.

The Arabinose Operon Gene Regulation. Why Gene Regulation? Developmental Changes Cell Specialization Adaptation to the environment Prevents creation of.

Yeast two hybrid screening of liver and WAT libraries I may need 15 instead of 10 minutes.

Transcription control elements (DNA sequences) are binding sites for transcription factors, proteins that regulate transcription from an associated.

Molecular Cell Biology of the Yeast Saccharomyces cerevisiae Lecture I: Biology, Genetics, Genomics and Proteomics Zhang Yi, National Institute of Biological.

GENETIC RECOMBINATION By Dr. Nessrin Ghazi AL-Abdallat Lecturer of Microbiology.

Lecture 9 Assignment 3 Yeast complementation analysis Discussion of Article 6.

Molecular Biology II Lecture 1 OrR. Restriction Endonuclease (sticky end)

Biol 729 – Proteome Bioinformatics Dr M. J. Fisher - Protein: Protein Interactions.

Restriction Enzyme Vector Ligase Enzyme Recombinant DNA DNA Construct Digestion ligation.

How many interactions are there? ~6,200 genes ~6,200 proteins x 2-10 interactions/protein ~12, ,000 interactions Yeast.

Molecular Cloning.

Protein-protein interactions Why study protein interactions? To infer function To understand regulatory networks Approach With given bait, discover target.

1 Genomics Advances in 1990 ’ s Gene –Expressed sequence tag (EST) –Sequence database Information –Public accessible –Browser-based, user-friendly bioinformatics.

The two-hybrid system – why?

Recombinant DNA Technology Studying protein interactions: Two Hybrid in yeast.

BioBuilding: What a Colorful World. Griffith and Bacterial Transformation.

By Rachel Schaefer Mentored by Mark Gomelsky and Min-Hyung Ryu Funded by Wyoming NSF EPSCoR.

YEAST MOLECULAR GENETICS (B)

Topics to be covers Basic features present on plasmids

Cloning Club Introducing a deletion into a DNA sequence -Khushbu

Yeast Two-Hybrid Screening: The Principles

Week 1: Tutorial Outline

pARA-R Sequence RFP Expression Sequence Biotechnology Lab Program

Biology II Study Guide for Unit Test on Operons and Transformation Lab 2013 You should be able to … 1. describe gene expression by explaining the following:

The Yeast Two-Hybrid System

Presentation transcript:

ACADEMIA SINICA IBMS 嚴仲陽美國德州貝勒醫學院博士 Specialty: Molecular & Cell Biology Hematopoiesis Apoptosis Regulation

Method Yeast Two-hybrid

Material Yeast strain：L40 (-his、-trp、-leu、-ade) Plasmid：pBTM116 ＆ pACT2 Competent Cell：HB101 Yeast DNA purification kit E coli. DNA purification kit

Result No Result ……

Normal Transcription

Yeast Two-hybrid Transcription

Plasmid construction

Bait ＆ Prey

Bait－pBTM116

Prey－pACT2

Yeast Two-Hybrid

Transformation ＆ Assay -Trp, -Leu, -His

β-galactosidase assay

gs3955 ＆ library Yeast Transformation Pick colonies Selection three times β-galactosidase assay Yeast DNA purification Transform to HB101 Bgl Ⅱ check Retransform to yeast Sequencing

My summer vacation Experiment Travel Camp