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Change in genetic structure of populations due to selective breeding by humans.
Limited by the genes available
using lving organisms or other biological systems to manufacture products.
Use genetic engineering
Recombinant DNA Technology:
Techniques used to alter the genetic material of an organism by adding new DNA fragments or altering existing DNA
Using recombinant DNA tecchnology to insert DNA from one organims to another resulting in a:
Recombinant DNA and Genetic Engineering:
Stages of genetic engineering:
1.) Find the gene of interest.
2.) Isolate the gene of interest
3.) Produce recombinant plasmid
4.) Get bacteria to take up plasmid
5.) Screen for clone of interest
Step 1. Find the gene of interest:
Know the sequeince of the gene
Know the lenght between restriction sites that surround the gene of interest
Restriction site: a particular base sequence found on DNA
a particular base sequence found on DNA
Step 2: Isolate the gene of interest
Cut DNA with a restriction endonuclease
Restriction endonuclease: an enzyme that cleaves DNA at a restriction site, usually within or near palindrome sequence.
1.) know sequence - use probe that has a radioactive or fluorescnet tag.
2.) know length -use gel electophoresis
an enzyme that cleaves DNA at a restriction site, usually within or near a palindrome sequence.
3.) Produce a recombinant plasmid
Vector: carriers recombinant DNA into a host cell
Plasmid: a small fragment of extrachromosal DNA, usually circular, that replicates independently of the main chromosome.
1.) Origin of replication site
2.) Marger gene (antibiotic resistant)
3.) Promoter / Terminator regions
-Cut plasmid w the same restriciton endonuclease
-Ligate to create recombinant plasmid
Step 4: Get bacteria to take up plasmid
The uptake of DNA directly from the environment into bacterial cell.
1.) increase membrane permeability
2.) mix bacteria w the vector
3.) further increase permeabilty w heat and salt
4.) allow bacteria to replicate
Why E. Coli?
Rapid growth and reproduction
Increase membrane permeability: Incubate w/ CaCl2
-bacteria membrane and plasmid both negatively charged.
-CaCL2 cover bacteria membrane with Ca++ and negatively charged plasimd is attracted to the cell
-Heat shock (produces pores)
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