Question 2 You wake up one morning to your roommate exclaiming about her sudden hair growth. She has been supplementing her diet with a strange new fungus purchased at the local farmers market. You take samples of the fungus to your lab and you find that this fungus does indeed make a protein(the harE protein) that stimulates hair growth. You construct a fungal genomic DNA library in the hope of cloning the hare gene. If you succeed you will be a billionaire! You obtain DNA from the fungus, digest it with a restriction enzyme, and clone it into a vector ampicillin resistance gene site for unwinding by helicase origin of replIcaton cloning a)Circle the vector that has the MINIMUM features required for your library construction. b)You clone your digested genomic DNA into this vector. What type of E. coli(bacteria)cells do you need to transform to create your library? You need ampicillin sensitive cells c)How do you distinguish bacterial cells that carry a vector from those that do not? Bacterial cells that carry a vector will be able to grow on ampicillin whereas untransformed cells will d)Circle on the following lists ALL you would need in order to construct the genomic DNA library. Assume you start with intact genomic dnA R Restriction enzyme CSize separation geD Ligase Okasaki fragments DNA Polymerase ATP TTP CTPGTP RNA Polymerase ddatp ddttp. ddctp Transcriptase ddGTP Reverse transcriptase 3 to 5 exonuclease Replication fork <Cloning vector Human cells 7012Fall20037.012 Fall 2003 3 Question 2 You wake up one morning to your roommate exclaiming about her sudden hair growth. She has been supplementing her diet with a strange new fungus purchased at the local farmer’s market. You take samples of the fungus to your lab and you find that this fungus does indeed make a protein (the harE protein) that stimulates hair growth. You construct a fungal genomic DNA library in the hope of cloning the harE gene. If you succeed you will be a billionaire! You obtain DNA from the fungus, digest it with a restriction enzyme, and clone it into a vector. a) Circle the vector that has the MINIMUM features required for your library construction. b) You clone your digested genomic DNA into this vector. What type of E. coli (bacteria) cells do you need to transform to create your library? You need ampicillin sensitive cells. c) How do you distinguish bacterial cells that carry a vector from those that do not? Bacterial cells that carry a vector will be able to grow on ampicillin whereas untransformed cells will not. d) Circle on the following lists ALL you would need in order to construct the genomic DNA library. Assume you start with intact genomic DNA. Enzymes Restriction enzyme Ligase DNA Polymerase RNA Polymerase Transcriptase Reverse Transcriptase 3' to 5' exonuclease Cloning vector Reagents Size separation gel Okasaki fragments ATP, TTP,CTP,GTP ddATP, ddTTP, ddCTP, ddGTP Primers Replication fork Human cells Virus = site for unwinding by helicase = origin of replication = promoter = cloning site = ampicillin resistance gene