02 Markers for Genetic Engineering

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02 Markers for Genetic Engineering

  1. 1. Markers for genetic engineering ALBIO9700/2006JK
  2. 2. Why fluorescent markers (easily stainedsubstance) are now used instead of antibiotic resistance markers• Antibiotic resistance markers – Original selected plasmid has antibiotic resistance genes to two different antibiotics (A and B) – Restriction enzyme is selected so that it cuts in the middle of one of these antibiotic resistance genes – Bacteria that have taken up the plasmid all have a successfully working copy of antibiotic resistance gene A. – Many plasmids also have a working copy of antibiotic resistance gene B, showing that the plasmids have failed to form recombinant DNA. – Bacteria that have taken up recombinant plasmids containing the cDNA insulin gene do not contain a working copy of the antibiotic resistance gene B. ALBIO9700/2006JK
  3. 3. ALBIO9700/2006JK
  4. 4. • Potential problem: plasmids commonly transferred between bacteria of the same species and also of different species meaning that pathogenic strains could receive plasmid with antibiotic resistance gene• Risk is a hypothetical one• Alternative method: – Incorporate a markergene for a protein that fluoresces green under ultra-violet light, along with desired genes (genes added using a micro-projectile to shoot them into plant cell nuclei) – quicker, higher proportion of transformed plants – Incorporate alongside the desired gene another marker gene that produces harmless product that is easily stained and is not normally produced by the cells ALBIO9700/2006JK
  5. 5. ALBIO9700/2006JK
  6. 6. ALBIO9700/2006JK

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