숙명여자대학교 / 생명과학과 박종훈 Biotechnology 의 최신 동향. 1. Genome: > 75 yrs 2. Genomics : T. Roderick in 1986 Structural genomics Functional genomics Functional Genomics.

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Presentation transcript:

숙명여자대학교 / 생명과학과 박종훈 Biotechnology 의 최신 동향

1. Genome: > 75 yrs 2. Genomics : T. Roderick in 1986 Structural genomics Functional genomics Functional Genomics

Post-Genome Era Unknown Genes Known Genes Functional Genomics cDNA Microarray Proteomics Bioinformatics Finding of novel functional genes Disease Drug target Human Genes  36,655

In vovo functional genomics?

Germline modification Isolate Blastocyst Derive ES cell line Gene Targeting Inject into Blastocyst Transfer to surrogate female Chimera Host blastocyst derived progenyES cell-derived progeny

Polycystic Kidney Disease (PKD) Autosomal Dominant Polycystic Kidney Disease (ADPKD) PKD1 - chromosome 16p 13.3 PKD2 - chromosome 4p21-q23 Non-PKD1-PKD2 PKHD1 - chromosome 6p 21.1-p12 Autosomal Recessive Polycystic Kidney Disease (ARPKD)

Structure of PKD1

Structure of PKD2

Characterization of mouse pkd2 and human PKD2 gene pkd1 PKD2 Physical spanning: ~42 kb 68 kb Transcript Size: 5.2 kb 5.4 kb ORF: ~2.9 kb 2.9 kb Amino Acids: Identical at Nucleotide level: ~87% Homologous at AA level: 90.8% Functional Domain: conserved Location: Chr. 5 Chr.4q21-23 Expression Level: Five fold greater in High in human ovary, mouse kidney than fetal and adult kidney, other organs testis, fetal lung, and intestine

Targeting of the Pkd2 Locus

The Pkd2 Null Mutation

EF-hand Domain

Primers for Mutagenesis Constructs of PKD2 C-terminal region 6th TM PGW2 B8-1 PGW3 PGW4 PGW5 EF-hand PKD2

-X-Z 6th TM B8-1 EF-hand PKD2 PGW6 -X-Z A B GST B8-1(W) B8-1(M) M GST PGW6(W) PGW6(-Z) PGW6(-X) C

What is cDNA microarray?

Sample Preparation Biological Question Microarray Life Cycle Microarray Reaction Microarray Detection Data Analysis & Modeling

DNA Chip 이란 무엇인가 ? 생명과학 + 전자, 기계공학 핵심기술 : 1) DNA Mcroarray 2) Biochip 3) Lab on a Chip DNA Chip

GeneFilters Microarrays a. Spotting Format b. Control c. Induced Sample

Selected DNA Microarray Photochemical technologyInk-Jet technologyGlass Slide

1. Gene Expression Analysis

Technology Access Target preparation Probe preparaion Data analysis

cDNA Microarray Analysis of DNA copy-number changes a b c

2. Affymetrix Chips

GeneChip Probe Array Synthesis Process 1. Light 2. Deprotected area Activation 3. Chemical coupling 4. New Mask 5. Coupling 6. Repeating PROCESS :

Sequence Analysis Array

Genotyping Arrays

Maskless Array DNA Chip Technology UV Light Source Filter Digital Micromirror Array Condenser From Computer From DNA Synthesizer Projection System DNA Chip being written

3. Nanogen DNA Chip Semiconductor Microchip Permeation Layer

Electronic Addressing of Nanogen DNA Chip

4. Hightlights of Clinical Micro Sensors DNA Chip CMS Technology Handheld DNA Chip Reader