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Ch. 22. Experimental Systems In vivo vs. in vitro systems Experimental animals

Ch. 22. Experimental Systems In vivo vs. in vitro systems Experimental animals Inbred strains reduce variation Adoptive transfer expts. p. 546. p. 547. Cell culture systems Primary lymphoid cell cultures are derived from blood or lymphoid organs.

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Ch. 22. Experimental Systems In vivo vs. in vitro systems Experimental animals

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  1. Ch. 22. Experimental Systems In vivo vs. in vitro systems Experimental animals Inbred strains reduce variation Adoptive transfer expts. Ch. 22

  2. p. 546 Ch. 22

  3. p. 547 Ch. 22

  4. Cell culture systems Primary lymphoid cell cultures are derived from blood or lymphoid organs. Cloned lymphoid cell lines are important tools. Hybrid lymphoid cell lines Monoclonal Ab’s T cell hybridomas, representing Th and Tc lineages Ch. 22

  5. p. 548 Ch. 22

  6. Ch. 22

  7. p. 550 Ch. 22

  8. Ch. 22

  9. Ch. 22

  10. Protein biochemistry Radiolabeling techniques - sensitivity Biotin labels combine with avidin Gel electrophoresis – proteins separated by size and charge X-ray crystallography – structural information Ch. 22

  11. p. 551 Ch. 22

  12. p. 552 Ch. 22

  13. Ch. 22 p. 552

  14. Ch. 22

  15. Ch. 22

  16. p. 553 Ch. 22

  17. p. 555 Ch. 22

  18. Ch. 22

  19. Recombinant DNA technology Restriction enzymes cleave DNA at precise sequences. DNA sequences are cloned into vectors, wherein DNA is replicated. Some vectors: bacterial and insect viruses, retroviruses, plasmids DNA clones are selected by hybridization. Ch. 22

  20. p. 556 Ch. 22

  21. p. 557 Ch. 22

  22. Ch. 22

  23. Ch. 22

  24. p. 557 Ch. 22

  25. Ch. 22

  26. Ch. 22

  27. p. 558 Ch. 22

  28. Southern blotting detects DNA of a given sequence. Northern blotting detects mRNA. The polymerase chain reaction (PCR) amplifies small amounts of DNA. Ch. 22

  29. p. 558 Ch. 22

  30. p. 559 Ch. 22

  31. Ch. 22 p. 560

  32. Gene transfer into mammalian cells Cloned genes transferred into cultured cells allow in vitro analysis of gene function. Cloned genes transferred into mouse embryos allow in vivo analysis of gene function. In knockout mice, targeted genes are disrupted. “Knock-in” technology allows the replacement of an endogenous gene. Ch. 22

  33. p. 562 Ch. 22

  34. p. 563 Ch. 22

  35. Ch. 22

  36. Ch. 22

  37. Ch. 22 p. 564

  38. p. 565 Ch. 22

  39. Ch. 22

  40. p. 566 Ch. 22

  41. Microarrays – An approach for analyzing gene expression “Expression profiling” Some are composed of cDNA Some are composed of oligonucleotides (“oligos”) Good diagnostic tool for some diseases Ch. 22

  42. p. 569 Ch. 22

  43. p. 570 Ch. 22

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