13-3 cell transformation

1,531 views

Published on

Published in: Education, Technology
0 Comments
2 Likes
Statistics
Notes
  • Be the first to comment

No Downloads
Views
Total views
1,531
On SlideShare
0
From Embeds
0
Number of Embeds
2
Actions
Shares
0
Downloads
95
Comments
0
Likes
2
Embeds 0
No embeds

No notes for slide
  • Knock out particular genes
  • 13-3 cell transformation

    1. 1. 13-3 Cell Transformation (Changes in Cell’s DNA)
    2. 2. Review• Which enzyme is used as a scissor in genetic engineering ? Which one is used as glue? Sciessor: Restriction Enzyme Glue: DNA ligase• What’s the function of gel electrophoresis? Separate DNA fragments by their different size
    3. 3. 13-3 Cell transformation• Cell transformation= change the cell’s DNA• A cell takes DNA from outside. The external DNA becomes part of the cell’s DNA
    4. 4. 1) Transforming bacteria cells – Using plasmid • small circle of extra bacterial DNA, double strand • Foreign DNA can be inserted into plasmid – Forms recombinant plasmids – Plasmids used in genetic engineering are called vectors. – Can deliver DNA into another cell
    5. 5. Plasmid
    6. 6. 1) Transforming bacteria cells – Using plasmid • Plasmid has a origin site for replication • Plasmid has a genetic marker (usually a gene for resistance to antibiotics) • two types of plasmid integration into a host bacteria
    7. 7. Restriction enzyme DNA or cDNA Foreign DNA, plasmidcuts molecule fragments with DNA, and modificationof chromosomal DNA sticky ends enzymes are mixedor cDNA recombinant plasmids containing foreign DNASame enzyme cuts plasmid DNAsame sequence in with sticky endsplasmid DNA host cells containing recombinant plasmids
    8. 8. What if insertion eukaryotic DNAinto bacteria ?Will there be any problems ???
    9. 9. Transcription RNA polymerase mRNA splicingDNA Pre-RNA mRNA No introns Bacteria don’t know how to deal with introns USE cDNA!!!
    10. 10. Reverse transcription= Making cDNA mRNA transcript Reverse transcriptasecDNA -- A DNA reversetranscription from a mRNA–cDNA hybridmatured mRNA `“c” -- complementary single-stranded cDNA DNA polymerase double-stranded cDNA
    11. 11. 2) Transforming plant cells• Recombinant plasmid is transformed to plant – By using bacterium Agropbacterium tumefaciens – By gene gun injection
    12. 12. By using Agrobacterium Plasmids carrying the gene of interest is transferred to agrobacterium The agrobacterium inserts the foreign DNA into the plant cell to form transgenic cell
    13. 13. By using gene gun To coat metal particles with gene containing plasmids Gene gun injects the DNA covered particles into the plant cell
    14. 14. 3) Transforming animal cells• Animal cell – Inject into egg nucleus some enzymes may help to insert foreign DNA into chromosomes – Gene replacement ( gene “knockout”)

    ×