GeneticEngineeringPreparedhumangeneNext, we introduce the prepared HUMAN gene to the mixture.If all goes according to plan, the human gene will fit into thecut in the plasmid so that the green T'gene will no longerworkcorrectly
目录 Genetic Engineering Next, we introduce the prepared HUMAN gene to the mixture. If all goes according to plan, the human gene will fit into the cut in the plasmid so that the green ‘T’ gene will no longer work correctly. Prepared human gene
GeneticEngineeringIntactPCeneand'defective"TgeneNo Por TgenePandTGenesintactAs plasmids are extremely small, we can not tell by looking whichones have got the human gene in the right place. We need to usea ‘shotgun' approach and incubate thousands of plasmids withhundreds of bacterial cells目聚
目录 Genetic Engineering As plasmids are extremely small, we can not tell by looking which ones have got the human gene in the right place. We need to use a ‘shotgun’ approach and incubate thousands of plasmids with hundreds of bacterial cells. No P or T gene Intact P gene and ‘defective’ T gene P and T Genes intact
GeneticEngineeringCellwithPandTintactRequiredcellCell withneitherPorTSome cells will take up the recombinant plasmid, some will takeup original plasmids, others will take up no plasmids at all or oneswithout antibiotic resistance genes
目录 Genetic Engineering Some cells will take up the recombinant plasmid, some will take up original plasmids, others will take up no plasmids at all or ones without antibiotic resistance genes. Required cell Cell with P and T intact Cell with neither P or T
GeneticEngineeringAgar containingpenicillinColonies growing fromsingle cells that areresistant to penicillinAn agar plate containing Penicillin is used to allow only those cellswhich have taken up a suitable plasmid to survive and divide. Thesecells must have resistance to Penicillin目聚
目录 Genetic Engineering An agar plate containing Penicillin is used to allow only those cells which have taken up a suitable plasmid to survive and divide. These cells must have resistance to Penicillin Agar containing penicillin Colonies growing from single cells that are resistant to penicillin
GeneticEngineeringNext, these colonies are sub-cultured onto agar containingtetracycline. Only cells resistant to BOTH antibiotics will beable to grow. We are interested in those cells which can't growinthepresenceofTetracycline
目录 Genetic Engineering Next, these colonies are sub-cultured onto agar containing tetracycline. Only cells resistant to BOTH antibiotics will be able to grow. We are interested in those cells which can’t grow in the presence of Tetracycline