In the transformation experiment you performed last week, you generated colonies
ID: 279598 • Letter: I
Question
In the transformation experiment you performed last week, you generated colonies of E.coli cells carrying twe different individual cells were expressing different sets of proteins encoded by genes on those plasmids. The diagram template appropriate. As you can see, space is limited, and you could get really bogged down in the details, so yoa should limit your plasmids: pCK lacZ and pCK:gfp. Those colonies exhibited different phenotypes when grown on differeest media because the below shows four individual cells, each carrying a plasmid. Complete these diagrams by adding the relevant proteins, where scope to four proteins, (?-lacta nase, Laci, ? galactosidase and GFP ) and three smal molecules (ampi ilin lactose, and al The media composition is indicated at the top of each box. Indicate what the colony phenotype would be for the The bacillus shape in each box represents one cell, the thick line is that cell's envelope. The circle inside of each cell is the .Be consistent in how you represent the proteins, and the substrates. (Hint: Use different symbols or colors for each Don't include things like RNA polymerase or mRNA as those are implied in diagrams like these indicated E colil/plasmid combination. The colony phenotypes should be in the notes from your lab work plasmid transformed into the cells with the loci of the genes indicated protein, substrate and product) Add a key or legend to help decipher your diagrams. You might chose to sacrifice detail for clarity, .Which of the plasmid-encoded proteins will be produced will be different depending on the meda compositionI conditions preclude production of a particular protein, don't put it on your diagram (or draw an X through it). lloteins are synthesized in the cytoplasm. but not all proteins stay there. if one or more proteins is/are exported outside the cells, make sure that is on your diagram. The space outside the cell is the medium the cell is growing on Pa ls regulated. Your diagrams should reflect this regulation, and should include a representation of how that regulation occurs. Your diagrams will be judged on the following criteria: . Accuracy: Does you diagram match the results in lab? Does your diagram describe thoseresults? o o Clarity: Neatness counts. The diagrams should focus on conveying they information, and while decorative flourish can be appreciated, if it is inaccurate or obscures key pieces of information, it can be penalized. Brevity. This is a diagram, not an essay. Responses containing more than 100 words of prose will be penalized. o LB Agar +Ampicillin Colony Phenotype: LB Agar Ampicillin Lactose Colony Phenotype: locz LB Agar+ Ampicillin+ Lactose+ X-gal+ and UV light Colony Phenotype: LB Agar Ampicillin+X-gal+ and UV light Colony Phenotype:Explanation / Answer
1) in first diagram colonies will be white because repressor is bound to lac operator.
2) in second condition colonies will be white. repressor is inhibited by lactose so lac operon gets activated. no X gal is present so no blue color production.
3) in third condition colonies will be blue in color. repressor is inhibited by lactose so lac operon gets activated. X gal is present and degraded by lac Z gene product. due to GFP protein cell will fluorescent under UV light.
4) in forth condition colonies will be white because repressor is bound to lac operator. no Lac Z production so no X gal degradation hence white colonies. due to GFP protein cell will fluorescent under UV light.