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GENETIC ENGINEERING RECOMBINANT DNA TECHNOLOGY

GENETIC ENGINEERING RECOMBINANT DNA TECHNOLOGY. HISTORY. 1972-73 Paul Berg, Herbert Boyer, Stanley Cohen and others investigated the properties of a new class of enzymes that cut DNA molecules in a unique way: RESTRICTION ENZYMES. T A C G A A T T C T G C A T T.

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GENETIC ENGINEERING RECOMBINANT DNA TECHNOLOGY

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  1. GENETIC ENGINEERING RECOMBINANT DNA TECHNOLOGY

  2. HISTORY 1972-73 Paul Berg, Herbert Boyer, Stanley Cohen and others investigated the properties of a new class of enzymes that cut DNA molecules in a unique way: RESTRICTION ENZYMES

  3. T A C G A A T T C T G C A T T A T G C T T A A GA C G T A A DNA

  4. T A C G A A T T C T G C A T T A T G C T T A A G A C G T A A DNA RESTRICTION ENZYME EcoR1 Cuts the DNA at the sequence GAATTC

  5. DNA T A C G A A T T C T G C A T T A T G C T T A A G A C G T A A RESTRICTION ENZYME EcoR1 Cuts the DNA at the sequence GAATTC Note that the cut breaks hydrogen bonds..and some covalent bonds

  6. RESTRICTION ENZYME EcoR1 Cuts the DNA at the sequence GAATTC Note that the cut breaks hydrogen bonds..and some covalent bonds DNA T A C G A A T T C T G C A T T A T G C T T A A G A C G T A A The cut ends have complementary ends…. Called “Sticky Ends”

  7. Bacterium PLASMID DNA “chromosome”

  8. Bacterium PLASMID DNA “chromosome”

  9. Bacterium “chromosome” REMOVE PLASMID DNA

  10. Bacterium “chromosome” CUT PLASMID DNA WITH EcoR1

  11. Human DNA with a gene for Growth Hormone

  12. Human DNA with a gene for Growth Hormone CUT THE HUMAN DNA WITH EcoR1

  13. Human DNA with a gene for Growth Hormone CUT THE HUMAN DNA WITH EcoR1 PIECE OF HUMAN DNA WITH GENE FOR GROWTH HORMONE

  14. Recombinant DNA technology “chromosome” Bacterium PLASMID DNA CUT WITH EcoR1 Both pieces of DNA Have “Sticky Ends” PIECE OF HUMAN DNA WITH GENE FOR GROWTH HORMONE CUT WITH EcoR1

  15. T A C G A A T T C T G C A T T A T G C T T A A G A C G T A A DNA RESTRICTION ENZYME EcoR1 Cuts the DNA at the sequence GAATTC

  16. Cuts the DNA at the sequence GAATTC RESTRICTION ENZYME EcoR1 The cut ends have complementary ends…. Called “ Sticky Ends ” Human DNA T A C G A A T T C T G C A T T A T G C T T A A G A C G T A A Gene for hormone Bacterial Plasmid DNA T A C G A A T T C T G C A T T A T G C T T A A G A C G T A A

  17. Recombinant DNA technology “chromosome” Bacterium (+) PLASMID DNA CUT WITH EcoR1 PIECE OF HUMAN DNA WITH GENE FOR GROWTH HORMONE CUT WITH EcoR1

  18. FLAVR SAVR ? A KIND OF TOMATO(E)

  19. PROBLEM: TOMATOES IN THE WINTER IN THE MIDWEST ARE TERRIBLE !!!! NO FLAVOR NO RED COLOR

  20. PROBLEM: TOMATOES IN THE WINTER IN THE MIDWEST ARE TERRIBLE !!!! NO FLAVOR NO RED COLOR WHY?????

  21. A. The tomatoes are picked and shipped BEFORE they are RIPE. Q. Why ????

  22. A. Because RIPE TOMATOESare too SOFT to ship (Ketchup in a box). Ripe Tomato: 1. Genes for LYCOPENE (RED PIGMENT)-making enzymes are switched on. 2. Genes for enzymes that make acids and flavors are switched on. 3. Genes for CELL WALL-DEGRADING ENZYMES are switched on.

  23. Ripe Tomato: 1. Genes for LYCOPENE (RED PIGMENT)-making enzymes are switched on. 2. Genes for enzymes that make acids and flavors are switched on. 3. Genes for CELL WALL-DEGRADING ENZYMES are switched on. What if we could turn GENES 1 and 2 on… …..but leave Gene 3 switched off? We should get a RIPE (Red and full of flavor)….but FIRM tomato Could be shipped to Iowa in January

  24. HOW ??? ANTI-SENSE TECHNOLOGY REMEMBER TRANSCRIPTION ??????

  25. TOMATO DNA CELL WALL- ENZYME GENE LYCOPENE GENE FLAVOR GENE

  26. TOMATO DNA mRNA RNA mRNA CELL WALL- ENZYME GENE LYCOPENE GENE FLAVOR GENE When the TomatoRIPENS, ALL 3 genes are TRANSCRIBED THE mRNAs are TRANSLATED and proteins are made

  27. TOMATO DNA mRNA mRNA mRNA CELL WALL- ENZYME GENE LYCOPENE GENE FLAVOR GENE When the TomatoRIPENS, ALL 3 genes are TRANSCRIBED THE mRNAs are TRANSLATED and proteins are made

  28. ? mRNA mRNA mRNA CELL WALL- ENZYME GENE LYCOPENE GENE FLAVOR GENE When the TomatoRIPENS, ALL 3 genes are TRANSCRIBED THE mRNAs are TRANSLATED and proteins are made What if we could INACTIVATE the mRNA for the cell wall-digesting enzyme?

  29. mRNA DNA The mRNA is a single strand of RNA made from ONE SIDE OF THE DNA THE “SENSE SIDE”.

  30. mRNA SENSE ANTISENSE DNA WHAT IF WE MADE an mRNA (single strand of RNA) ... from FROM THE OTHER SIDE OF THE DNA THE “ANTI- SENSE SIDE” ?????.

  31. THE 2 RNAs ARE COMPLEMENTARY SENSE mRNAs ANTISENSE Sense mRNA Anti-Sense mRNA GENE GENE DNA WHAT IF WE INSERTED A NEW GENE, ONE WITH THE BASE SEQUENCE FROM THE OTHER SIDE OF THE DNA... THE “ANTI- SENSE SIDE” ?????.

  32. mRNA SENSE ANTISENSE mRNA THE 2 RNAs ARE COMPLEMENTARY WHEN THE mRNAs PAIR… THEY CAN NOT BE TRANSLATED Sense mRNA Anti-Sense mRNA GENE GENE DNA WHAT IF WE INSERTED A NEW GENE, ONE WITH THE BASE SEQUENCE FROM THE OTHER SIDE OF THE DNA... THE “ANTI- SENSE SIDE” ?????…AND WHAT IF WE TRANSCRIBED BOTH GENES AT THE SAME TIME???

  33. mRNA SENSE ANTISENSE mRNA THE 2 RNAs ARE COMPLEMENTARY WHEN THE mRNAs PAIR… THEY CAN NOT BE TRANSLATED DNA WHAT IF WE MADE an mRNA (single strand of RNA) ... from FROM THE OTHER SIDE OF THE DNA THE “ANTI- SENSE SIDE” ?????.

  34. TOMATO DNA Antisense mRNA mRNA mRNA mRNA CELL WALL ENZYME GENE COMPLEMENT (ANTISENSE) FLAVOR GENE LYCOPENE GENE CELL WALL- ENZYME GENE When the TomatoRIPENS, ALL 3 genes are TRANSCRIBED AND….the antisense mRNA is made for the enzymes that break down The tomato cell walls (make it soft).

  35. TOMATO DNA Antisense mRNA + mRNA mRNA mRNA CELL WALL ENZYME GENE BACKWARDS (ANTISENSE FLAVOR GENE LYCOPENE GENE CELL WALL- ENZYME GENE When the TomatoRIPENS, ALL 3 genes are TRANSCRIBED AND….the antisense mRNA is made for the enzymes that break down The tomato cell walls (make it soft).

  36. TOMATO DNA Antisense mRNA + mRNA mRNA mRNA CELL WALL ENZYME GENE BACKWARDS (ANTISENSE FLAVOR GENE LYCOPENE GENE CELL WALL- ENZYME GENE When the TomatoRIPENS, ALL 3 genes are TRANSCRIBED AND….the antisense mRNA is made for the enzymes that break down The tomato cell walls (make it soft).

  37. CORN: Insert a gene for the TOXIN (Bt)(protein) from Bacillus thuringiensis (a bacterium) Into Corn Bt toxin kills insects…Don’t need a Pestidcide Spray

  38. Soybeans Insert a gene for RESISTANCE TO “Roundup” Herbicide (Weed Killer) Plant seeds Germinate (soybeans and Weeds) Spray whole field with Roundup…. WEEDS DIE….SOYBEANS LIVE

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