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WSU / Biology / BIO 1510 / What are restriction enzymes used for?

What are restriction enzymes used for?

What are restriction enzymes used for?

Description

School: Wayne State University
Department: Biology
Course: Basic Life Mechanisms
Professor: Bradley thomas
Term: Fall 2016
Tags:
Cost: 50
Name: BIO 1050 Exam 4 study guide
Description: lecture echos and also my notes added
Uploaded: 03/30/2017
38 Pages 46 Views 3 Unlocks
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EXAM 4If you want to learn more check out What are the secondary hazards of earthquakes?

CHAPTER 12If you want to learn more check out What are the stages of the water cycle?

DNA Technology and Genomics:We also discuss several other topics like What areas of the brain are affected by adhd?
We also discuss several other topics like What is biosphere's description in short answer?

  • If you want to do research on DNA, you need to know how to cut DNA…
  • Humans have 25,000 encoding genes.
  • Reason why we cut DNA - it’s easier to work with smaller ones than larger ones.

  • Restriction enzymes- (shown in purple in ppt.) Group of enzymes that cut DNA at a specific site. EcoRI- type of enzyme that helps cut DNA at a specific site (restriction site). It cuts the restriction site. Always between G&A.

  • Sticky End- overhanging simple stranded end
  • Sticky ends of the bacterial DNA can bind to the sticky ends of human DNA as long as both bacterial and human DNAs are cut with the same restriction enzyme.
  • DNA ligase creates covalent bonds that joins a recombinant DNA

  • So basically…

  1. Every restriction enzyme recognizes one specific nucleotide sequence (its restriction site)
  2. A restriction enzyme always cuts DNA sequences at its restriction site in an identical manner.
  3. A piece of DNA from another source (the gene of interest) is cut by the source restriction enzyme.
  4. The DNA fragments from the two sources stick together by hydrogen bonding of the base pairs.
  5. The enzyme DNA Ligase creates new covalent bonds that join the backbones of the DNA strands. The result is a piece of the recombinant DNA.

If you want to learn more check out What is production possibility curve as explained with diagram?

  • Recombinant DNA- has nucleotide sequences belonging to different organisms. We need the same enzyme and DNA Ligase
  • What can you do with the DNA you cut up? (Make a few copies.)
  • In the gene of interest it encodes protein V, we want to make many copies.

Don't forget about the age old question of What are tetrahedral sigma bonds?

  • We need to introduce bacteria…
  • Plasmid= small circular double stranded DNA that replicates separately from the bacterial chromosome.
  • Plasmid can be removed from the bacteria without harming H

  • So basically…

  1. Plasmid is isolated
  2. The cell's DNA is isolated. (A cell with DNA containing the gene of interest)
  3. The plasmid is cut with an enzyme.
  4. The cell's DNA is cut with the same enzyme.
  5. The targeted fragment and plasmid DNA are combined.
  6. DNA Ligase is added, which joins the 2 DNA molecules.
  7. Which makes a recombinant DNA plasmid.
  8. The recombinant plasmid is taken up by a bacterium through transformation
  9. The bacterium is produced.

  1. Two things can occur:
  1. Harvested proteins may used directly; or
  2. Genes may be inserted into other organisms.

  • Now that we have copies of a gene, what can we do with it?

  • Recombinant DNA is produced which contains Gene V.
  • This is taken by bacteria known as transformation.

  • Recombinant bacterium then divides many times by binary fission, to produce many copies of Gene V.

  • Copies of Gene V are transcribed and translated to make many copies of protein V.

  • We can then insert gene V with other organisms making transgenic organisms.
  • Transgenic organisms contain genes from other organisms of other species.

  • Remember:
  • Without bacteria we can't transcribe or translate copie of gene V. We won't be able to make more copies.

  • Table 12.6

Do not worry about the table just know:

  • E. coli can be used for many things:
  • Insulin
  • Growth hormones

  • Using DNA to change a genome..
  • T1 plasmid- came from soil bacterium, Agrobacterium tumefaciens. This is a tumor-inducing plant.

  • Recombinant T1 plasmid- lacks tumor inducing gene, but has a new gene.

shown in red pictures

  • DNA containing the gene for a desired trait- gene encoding vitamin, pesticide resistance gene, or drought surviving gene.

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