Gene Expression

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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

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What Is Gene Expression?? (Protein Expression) 3 Gene Expression 1/22/2011

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Gene Expression = Gene + expression Gene : 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.” Expression : The process of expressing a gene.. The representation of characterstics or signs of gene.. 4 Gene Expression 1/22/2011

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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

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1/22/2011 Gene Expression 7 Protein Folding And Transport Synthesis of RNA from DNA by RNA polymerase Enzyme DNA  RNA Become mature RNA RNAm-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.

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1/22/2011 Gene Expression 8 Gene Expression : Diagnostic Tests

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1/22/2011 Gene Expression 9

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1/22/2011 Gene Expression 10 1.) Blood Test

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(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

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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

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1/22/2011 Gene Expression 18 Southern Blotting Technique Southern Blotting Study of DNA DNA is detected by Probe (DNA/RNA)

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1/22/2011 Gene Expression 19 Northern Blotting Technique Northern Blotting Study of RNA RNA is detected by Probe (DNA/RNA)

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1/22/2011 Gene Expression 20 Western Blotting Technique Western Blotting Study of Proteins Protein is detected by Antibody

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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

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1/22/2011 Gene Expression 25

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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

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1/22/2011 Gene Expression 28

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1/22/2011 Gene Expression 29 Gene Expression : Current Uses

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1/22/2011 Gene Expression 30 Current Uses

Plant Physiology : 

Plant Physiology Gene expression changes by microarray technique used to study plant physiology.. Eg: 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

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1/22/2011 Gene Expression 33 Gene Expression : Summary

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1/22/2011 Gene Expression 34 Gene Expression Defination : GeneDNARNAm-RNAProtein M/A : Transcription Translation Protein synthesis Diagnostic Tests : Blood Test Course CAD test DXd Test Micoarray based test Analytical Techniques : Reporter gene Blotting Techn DNA microarray SAGE Current Uses : Current Uses : To study Plant Physiology TNT Gene Transfer Technology In AML patient

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1/22/2011 Gene Expression 35