Recombinant DNA - PowerPoint PPT Presentation

1 / 9
About This Presentation
Title:

Recombinant DNA

Description:

Recombinant DNA Scope Human Genome = 3x109 Average Gene = 3x104 (1/105) SNP Mutation (1/109) Process Cut DNA into pieces Insert DNA into vectors that can replicate in ... – PowerPoint PPT presentation

Number of Views:65
Avg rating:3.0/5.0
Slides: 10
Provided by: dnaCsByu
Learn more at: http://dna.cs.byu.edu
Category:
Tags: dna | recombinant

less

Transcript and Presenter's Notes

Title: Recombinant DNA


1
Recombinant DNA
2
Scope
  • Human Genome 3x109
  • Average Gene 3x104 (1/105)
  • SNP Mutation (1/109)
  • Process
  • Cut DNA into pieces
  • Insert DNA into vectors that can replicate in
    bacteria
  • Transform (introduce) DNA into host cell
  • Plate cells and select those with vectors
  • Each colony has one chunk of DNA
  • The whole set is a library of human DNA

3
Specifics
  • Restriction enzyme GAATTC
  • Palindrome
  • Exists for a large number of loci
  • Ligase can bind together DNA from any organism
    that has been cut with the enzyme
  • Comes from bacteria that need to repair their own
    DNA
  • Hook fragment to bacterial DNA with a replication
    site.

4
Episomes
  • Contain resistance genes
  • Are transferred between bacteria to allow for
    more rapid adaptation
  • We can insert all of the fragments from the human
    genome into bacterial episomes and create a
    library of bacterial colonies that can be used to
    replicate DNA

5
Reverse Translation
  • You can take a protein like beta-globin and
    determine the amino acid sequence
  • You can then create a set of complimentary probes
    for all possible nucleotide sequences that would
    cause the amino acid sequence
  • Dump the probe on each entry in the library to
    find the gene that makes beta-globin

6
Other Approaches
  • Insert DNA into a virus and infect malalian
    cells.
  • Create artificial chromosomes and inject in the
    nucleus
  • Bacterial Artificial Chromosome
  • Stem Cell Manipulation

7
Lets grow insulin
  • Insert insulin gene into episome
  • But promoters for e-coli are different than human
  • So put e-coli promoter on the front of episome
  • But humans do splicing and bacteria dont
  • So, we need to insert spliced gene into epiisome
  • Use reverse transcriptase to turn RNA into DNA
  • Viruses use this to replicate
  • But protein may fold differently in bacteria, so
    you may have to do some more engineering

8
How about mammals
  • Rabies antibodies are important
  • It is hard to find people that generate them
  • People dont like to be infected to generate
    antibodies for pharma companies
  • Put human gene that generates antibody into
    mouse, infect it and collect antibodies
  • But mice dont have much blood, lets do it with a
    cow
  • But the cows immune system also generates cow
    antibodies
  • So lets cut out the cow immune system genes and
    totally replace them with human immune system
  • Hope that the cow doesnt get sick

9
(No Transcript)
Write a Comment
User Comments (0)
About PowerShow.com