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“ The Discovery of Reverse Transcriptase”

November 2, 2010 Biology 161 Dr. Hong Fei By: David Meyer Kylene Wolfe Ryan Schramer Adrian Bachman. “ The Discovery of Reverse Transcriptase”. History. Genetic information was demonstrated through the transfer of information of the biological system during the years of:

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“ The Discovery of Reverse Transcriptase”

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  1. November 2, 2010 Biology 161 Dr. Hong Fei By: David Meyer Kylene Wolfe Ryan Schramer Adrian Bachman “The Discovery of Reverse Transcriptase”

  2. History • Genetic information was demonstrated through the transfer of information of the biological system during the years of: • 1940's, 1950's & 1960's • In 1970 more of a focus on RNA tumor viruses but left many unanswered questions.

  3. Howard Temin • Studied RNA tumor viruses most of his life • His early days he spent time working on Rous sarcoma virus (RSV) • Temin was the one that came up with the first hypothesis for the study of Reverse Transcriptase

  4. “The RNA genome of RSV is converted into a DNA provirus.” Howard Temin Hypothesis

  5. “RSV is sensitive to inhibitors of DNA synthesis and suggesting that DNA, complementary to the RSV genomic RNA, is present in transformed cells.” Howard Temin Proposal

  6. David Baltimore • Spent time studying virus replication • He looked at RNA and DNA at a more biochemical view looking deep within individual virions • He later focused on RNA tumor viruses and concluded with Rauscher murine leukemia virus (R-MLV)

  7. Howard Temin and David Baltimore walked two different pathways

  8. They disrupted their pure stock viruses by using nonionic detergents “Can a retrovirus perform DNA synthesis?” Then “radiolabeled deoxthymidine triphosphate (dTTP) along with three deoxynucleotide triphosphates (dATP, dCTP, dGTP) to the virion preparation.” Experiment Time!

  9. YES! Each experiment, dTTP turned into nucleic acid Did these reactions contain any incorporation of radioactive dTTP into DNA?

  10. Results: Sensitive to Dnase They tested their experiment by adding enzymes reacted with RNA or DNA

  11. “What was the template?” One more question?

  12. Baltimore and Temin both “preincubated the virions with RNase, which catalyzes the degradation of RNA into ribonucleotide monophosphates.” One more test!

  13. “The longer the pretreatment of virions with RNase, the lower the amount of DNA-synthesizing activity, - thus an enzyme in the virion could catalyze RNA-dependent DNA synthesis.” Results:

  14. Demonstration of RNA-dependent DNA Synthesis Radioactive Tymidine Incorporated into Nucleic Acid Experimental Treatment: R-MLV RSV Standard Conditions 3.69 pmol 9110 dpm Virions pretreated with RNase 0.52 pmol 2650 dpm Untreated Product 1425 dpm 8350 dpm After product is treated with RNase 1361 dpm 7200 dpm After Product is treated with Dnase 125 dpm 1520 dpm Results/Data

  15. “Reverse Transcriptase” This was reverse to what normally was happened in DNA and RNA giving it's name:

  16. Temin believed that his hypothesis was true all along he just had to prove it to the world Baltimore believed a different theory but tested Temin theory by checking RNA-dependent and DNA polymerization. Conclusion:

  17. Concert mRNA to DNA help create cDNA libraries coloning and study of tissues study of many genes throughout biology Explosion of research in retroviruses/ RNA viruses What has this done to help the world?

  18. Temin/Baltimore Future Since HIV-AIDS is a retrovirus! This drove Temin and Baltimore to a Noble Prize in 1975

  19. 1. Why was the research done? a. Win a Noble prize b. Become president c. To test whether or not RNA genome of RSV is converted into a DNA provirus d. Test Michael Jackson's DNA QUIZ TIME!!!

  20. 2. What method did these men use? a. Newton's Law method b. Broke down a virus and started doing DNA/RNA test on it c. Dr. Fei Method d. Heartland College Education Method Question 2

  21. 3. With the results was this experiment successful? a. YES! b. No c. Maybe d. Not sure Question 3

  22. 4. What could you conclude from this experiment? a. It was the worlds greatest experiment b. The longer the pretreatment of virions with RNase, the lower the amount of DNA-synthesizing activity, - thus an enzyme in the virion could catalyze RNA-dependent DNA synthesis.” c. If water goes up, it must come back down d. DNA/RNA is the same in all people Question 4

  23. 5. What is one way this research has helped the world? a. It helped two men win a Noble Prize b. It's going to save the world c. It gave a cure to AIDS d. Explosion of research in retroviruses/ RNA viruses Question 5

  24. Who is the greatest Biology instructor in the who world? a. Dr. Hong Fei b. President Obama c. Rod Blagojevich d. Police Officer Bonus Question

  25. The End!

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