Slide 1: Gene Expression Compiled by,
Juhi Shah 1 Gene Expression 1/22/2011 Outline : Outline Gene Expression : Defination
Gene Expression : MOA
Gene Expression : Diagnosis
Gene Expression : Analytical Techniques
Gene Expression : Current Uses
Gene Expression : Summary 2 Gene Expression 1/22/2011 Slide 3: What Is
(Protein Expression) 3 Gene Expression 1/22/2011 Slide 4: Gene Expression = Gene + expression
It is segment of DNA..
It is passed from parents to a child..
Genes determine the child's characteristics – like : hair, eye, skin color,height – and whether the child will have an inherited disease.”
The process of expressing a gene..
The representation of characterstics or signs of gene.. 4 Gene Expression 1/22/2011 Slide 5: Gene Expression :
“Process by which a gene's information is converted into the structures and functions of a cell. “
It measure the activity (the expression) of thousands of genes.
genes are first converted to DNA--> RNA--> m-RNA-->Proteins 5 Gene Expression 1/22/2011 Gene Expression M/A : 1/22/2011 Gene Expression 6 Gene Expression M/A Slide 7: 1/22/2011 Gene Expression 7 Protein Folding And Transport Synthesis of RNA from
DNA by RNA polymerase Enzyme
DNA RNA Become mature RNA
RNAm-RNA m RNA Protein Protein Folding
Chaperone enz involved .
Form 3-D struct of protein Protein Transport
Transport of proteins is occur to the endoplasmatic reticulum, transport via the Golgi apparatus. Slide 8: 1/22/2011 Gene Expression 8 Gene Expression :
Diagnostic Tests Slide 9: 1/22/2011 Gene Expression 9 Slide 10: 1/22/2011 Gene Expression 10 1.) Blood Test (Cont.) : (Cont.) Heritable factors : derived from parents generate gene expression changes
External factors : air pollution /cigarette smoke generate gene expression changes in the body.
Internal factors : colon tumor,precipitate changes in gene expression in blood cells
Blood is taken RNA extracted snapshot of specific gene expression changes derived from microarray anaylsis the disease snapshot and normal snapshot are clearly different. 1/22/2011 Gene Expression 11 2.) Course CAD Test : Measure the amounts of specific types of RNA which produce atherosclerosis.
The test provides information about the current health of arteries. 1/22/2011 Gene Expression 12 2.) Course CAD Test 3.) XDx Test XDx is a molecular diagnostics company.
It is non-invasive gene expression testing
during Heart Transplant Management patients Slide 13: 1/22/2011 Gene Expression 13 Gene Expression : Analytical Techniques Techniques : Techniques 1/22/2011 Gene Expression 14 1.) Reporter Gene : 1.) Reporter Gene It act as a reporter..
It is used to analyze another gene activity.
It determine whether a particular DNA construct has been successfully introduced into a plant cell/tissues or animal organs. 15 Gene Expression 1/22/2011 2.) Blotting Techniques : Transferring DNA,RNA and proteins onto a carrier so they can be separated.. : 2.) Blotting Techniques : Transferring DNA,RNA and proteins onto a carrier so they can be separated.. 16 Gene Expression 1/22/2011 Steps of Blotting Technique : Steps of Blotting Technique Run sample (DNA/RNA/Proteins) on a gel
Transfer it to a membrane
Link sample to membrane
Make a probe (DNA/RNA/Antibody)
Hybridize probe to membrane
Detect probe 1/22/2011 Gene Expression 17 Slide 18: 1/22/2011 Gene Expression 18 Southern Blotting
Technique Southern Blotting
Study of DNA
DNA is detected by
Probe (DNA/RNA) Slide 19: 1/22/2011 Gene Expression 19 Northern Blotting
Technique Northern Blotting
Study of RNA
RNA is detected by
Probe (DNA/RNA) Slide 20: 1/22/2011 Gene Expression 20 Western Blotting Technique Western Blotting
Study of Proteins
Protein is detected by
Antibody Slide 21: Gene Probe :
It is fragment of DNA or RNA marked by a chemical or radioactive substance..
It will bind with gene and identify or isolate that gene.. 1/22/2011 Gene Expression 21 3) RT – PCR : (Formation of DNA from RNA) : 3) RT – PCR : (Formation of DNA from RNA) Deoxy ribonucleotide tri phoshphate 4.) DNA Microarray : 4.) DNA Microarray Also known as DNA Chip/ Gene chip
It measure the transcription level for every gene in a gene expression
(Transcription : DNA RNA mRNA)
Microarray detects mRNA 1/22/2011 Gene Expression 23 A small 1 square centimeter chip that’s divided into thousands of squares.
Each square contains many copies of a single gene. Applications of DNA microarray : Applications of DNA microarray To identify mutation in a gene..
Analyze gene activity changes in sample over longer period of time..
Compare gene expression (genotype) in different samples..
(eg. Gene expression changes in normal cell and tumour cell) 1/22/2011 Gene Expression 24 Slide 25: 1/22/2011 Gene Expression 25 Slide 26: 1/22/2011 Gene Expression 26 5.) SAGE : 5.) SAGE SAGE : “Serial Analysis of Gene Expression.”
SAGE is a method for: analysis of gene expression patterns.
3 Principles of SAGE :
A short sequence of DNA tag (10-14bp),contains information to identify a transcription
Sequence tags can be linked together;
Quantitate tags and determine patterns of gene expression. 1/22/2011 Gene Expression 27 Slide 28: 1/22/2011 Gene Expression 28 Slide 29: 1/22/2011 Gene Expression 29 Gene Expression :
Current Uses Slide 30: 1/22/2011 Gene Expression 30 Current Uses Plant Physiology : Plant Physiology Gene expression changes by microarray technique used to study plant physiology..
Cotton mutants = due to altered fibre characterstics.
Elongation of maize root = due to low soil water potential. 1/22/2011 Gene Expression 31 TNT (Tissue Necrosis Therapy) Lonza use the GS (glutamine synthatase) gene expression system for the :
manufacture,therapeutic and diagnostic products based on the TNT (Tissue Necrosis Therapy) which generate chimeric and fully human TNT antibodies. Gene Transfer Technology : Gene Transfer Technology Gene transfer technology to treat AIDS,cardiopathies, and neurologic diseases..
In AML Patients
c-DNA microarrays to determine the levels of gene expression in blood samples or bone marrow samples 1/22/2011 Gene Expression 32 Slide 33: 1/22/2011 Gene Expression 33 Gene Expression : Summary Slide 34: 1/22/2011 Gene Expression 34 Gene Expression Defination :
GeneDNARNAm-RNAProtein M/A : Transcription
Protein synthesis Diagnostic Tests : Blood Test
Course CAD test
Micoarray based test Analytical Techniques : Reporter gene
SAGE Current Uses : Current Uses :
To study Plant Physiology
Gene Transfer Technology
In AML patient Slide 35: 1/22/2011 Gene Expression 35