Bacterial Transformation – bacteria take up and express foreign DNA, usually a plasmid. Plasmid – circular piece of DNA.

Slides:



Advertisements
Similar presentations
Transformation and Cloning
Advertisements

Lecture 8 Genetic Engineering. Medically important substances produced by genetic engineering Human Insulin- used to treat diabetes Past: extracted insulin.
Introduction of DNA into Living Cells
Gene technology - what is it? - what is it used for? - how does it work?
Recombinant DNA Technology “Gene Cloning”. What is it?  Gene cloning: production of large quantities of a specific, desired gene or section of DNA to.
PGLO™ Transformation. Central Framework of Molecular Biology DNA RNA ProteinTrait.
Ability to replicate independently (so that a lot of copies could be generated) A recognition sequence for a restriction enzyme (so that we can introduce.
Bacterial Transformation. Broad and Long Term Objective To characterize a single clone from an Emiliania huxleyi cDNA library using sequence analysis.
Genetic engineering Recombinant DNA technology. Questions: Name 3 things you know about bacteria. What are some characteristics that make bacteria a good.
Transfection The students need to have some background knowledge about recombinant DNA technology for this lecture. Key words: Transient transfection,
CHAPTER 4 DNA CLONING (cont.) MISS NUR SHALENA SOFIAN.
2nd lab competent cells formation and transformation of competent cells with DNA. BCH 462 [practical]
PGLO™ & GFP.
Competent cells formation and transformation of competent cells with DNA. BCH 462 [practical] 2 nd lab.
Bacteria Transformation
Lab Exam 2 Overview. Bacterial Transformation To impart new phenotype by adding expressible genes Why use bacteria? – Rapid growth – Plasmids as vectors.
© SSER Ltd..
Plasmid DNA Restriction Enzymes “cut” Plasmid DNA Piece of DNA is Removed New Piece (gene) of DNA is “stitched” to Plasmid DNA New DNA (gene)
GENETIC ENGINEERING (RECOMBINANT DNA TECHNOLOGY)
In vivo gene cloning. Can you remember... What we mean by in vitro and in vivo?
In vivo gene cloning.
Genetic Engineering. “In Greek mythology, the chimera was part lion, part goat, part serpent, which was slain by the hero Bellerephon. In modern day biology,
Making Recombinant DNA DNA structure and Plasmids DNA Restriction and Ligation
13–2Manipulating DNA A.The Tools of Molecular Biology 1.DNA Extraction Homogenization: Cell walls, membranes, and nuclear material are broken Emulsification:
Bacterial Transformation
Introduction to pGLO lab Bacteria Transformation Please take these notes carefully. You do not need to write anything in RED.
Chapter 13 Genetic Engineering
PGLO ™ & GFP. Central Framework of Molecular Biology DNA RNA ProteinTrait.
Cloning Genes Gene cloning: amplifying a specific piece of DNA via a bacteria cell Cloning vector: a replicating DNA molecule attached with a foreign DNA.
Transformation of E.coli with pGal. Exchange of Genetic Information in Bacteria 1.Transformation 2.Transduction 3.Conjugation.
Biotechnology Techniques
Biotechnology Practice Test. Question #1 An organism’s chromosomes are part of its a) plasmid b) recombinant DNA c) genome d) enzymes.
Fall Electrophoresis is a molecular technique that separates nucleic acids and proteins based on Size and +-+ Charge +-+
Fall Electrophoresis is a molecular technique that separates nucleic acids and proteins based on Size and +-+ Charge +-+
Bacterial Transformation
DNA Science. Restriction Digest Restriction Digestion is the process of cutting DNA molecules into smaller pieces with special enzymes called Restriction.
Restriction Enzyme Vector Ligase Enzyme Recombinant DNA DNA Construct Digestion ligation.
DNA Technology Part 2.
Plasmids and Vectors Aims:
+ genetic engineering module 2 – biotechnology & gene technologies.
Transformation Lab What are plasmids? Circular sequences of DNA that can be incorporated into a bacterial host genome. What makes them so special?? They.
Bacterial Transformation
Transformation MISS : SALSABEEL H. AL- JOUJOU.
Bacterial Transformation Green Fluorescent Protein.
Recombinant DNA Plasmids and Bacteria Transformation.
Transformation Biology experiment. Historical Perspective Frederic Griffith 1928 London First controlled demonstration of genetic transformation Griffith.
DNA molecules from 2 different species, if cleaved by the same
Lab# 2 Competent Cells Formation and Transformation of Competent Cells with plasmid DNA. BCH 462 [practical]
Bacterial Transformation – bacteria take up and express foreign DNA, usually a plasmid. Plasmid – circular piece of DNA.
V Cell Transformation Recombinant DNA Host Cell DNA Target gene
Biotechnology Practice Test
Genetic Research and Biotechnology Recombinant technology
Methods of transformation
Real Cats Wear Pink Lab Process Explanation
DNA Technology Part 2.
© SSER Ltd..
13-3 Cell Transformation Interactive pgs. 329.
Genetic Research and Biotechnology Recombinant technology
Genetic Engineering Insulin production Extra-nutrient foods
Chapter 13.3 Cell Transformation.
Lab# 2 Competent Cells Formation and Transformation of Competent Cells with plasmid DNA. BCH 462 [practical]
Copyright Pearson Prentice Hall
Introduction to the pGLO Lab
What is Transformation?
The Genetics of Bacteria
Copyright Pearson Prentice Hall
Cell Transformation.
Bacterial Transformation
DEPARTMENT OF MICROBIOLOGY, S.M. JOSHI COLLEGE, HADAPSAR, PUNE
Plasmids circular pieces of”extrachromosomal” DNA propagated inside host have origin of replication -> ensures host will copy it.
Presentation transcript:

Bacterial Transformation – bacteria take up and express foreign DNA, usually a plasmid. Plasmid – circular piece of DNA

Steps of Bacterial Transformation Choose a bacterial host. E.coli is a model organism. Well studied No nuclear membranes Has enzymes necessary for replication DNA/ Chrom.

Steps of Bacterial Transformation 2. Choose a plasmid to transform. Characteristics of a useful plasmid. Single recognition site Plasmid only cuts in one place, so this ensures that the plasmid is reformed in the correct order. Origin of replication Allows plasmid to replicate and make copies for new cells. Marker genes Identifies cells that have been transformed.  gene for antibiotic resistance – bacteria is plated on media with an antibiotic, and only bacteria that have taken up a plasmid will grow  gene that expresses color – bacteria that have taken up a recombinant plasmid are a different color than bacteria that have taken up a NONrecombinat vector.

Steps of Bacterial Transformation 3. Prepare bacterial cells for transformation of plasmid. Treat with calcium chloride, which allows plasmid to pass through bacterial cell walls. This is the most common method. Electroporation - brief electric pulse Directly inject plasmid into bacterial cell.

Steps of Bacterial Transformation 4. Plate transformation solution on appropriate media. Contains nutrients for bacteria. Contains antibiotic to distinguish transformed bacteria from NONtransformed bacteria. 5. Incubate plates overnight. E.coli grows in the human body, and is therefore incubated at body temperature (37°C) 6. Analyze plates.