Your colleague handed you a novel strain of coli that is purifying a protein with a 6xHisTag; they claim it is superior to the TOP10 cells you have been using. But even with a larger culture size, you discover that your protein yield—using the same Ni-NTA column—is quite low. You discover that the novel strain of Escherichia coli generates an unusually high quantity of dicarboxylic acids, a byproduct of the citric acid cycle that is recognized for its ability to function as an all-purpose metal chelator. What do you think the issue is with purifying IMAC protein with this new strain of E. coli?
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Your colleague handed you a novel strain of coli that is purifying a protein with a 6xHisTag; they claim it is superior to the TOP10 cells you have been using. But even with a larger culture size, you discover that your protein yield—using the same Ni-NTA column—is quite low. You discover that the novel strain of Escherichia coli generates an unusually high quantity of dicarboxylic acids, a byproduct of the citric acid cycle that is recognized for its ability to function as an all-purpose metal chelator. What do you think the issue is with purifying IMAC protein with this new strain of E. coli?
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- In a nutrient medium that lacks histidine, a thin layer of agar containing ~109 Salmonella typhimurium histidine auxotrophs (mutant cells that require histidine to survive) produces ~13 colonies over a two-day incubation period at 37 °C. How do these colonies arise in the absence of histidine? The experiment is repeated in the presence of 0.4 μg of 2-aminoanthracene. The number of colonies produced over two days exceeds 10,000.What does this indicate about 2-aminoanthracene? What can you surmise about its carcinogenicity?in isolating ribosomes from a yield sample, describe the ideal type of centrifugation for this separation technique based on the following:*Main type of centrifuge *Subtype centrifuge * Speed range *Operational temperature range*Type of CentrifugationTRUE OF FALSE: Is E coli colonies would be pink mucoid in Eosin Methylene BlueAgar (EMBA)? Is Coliform colonies would be pink mucoid in Eosin Methylene BlueAgar (EMBA)?
- You have several different media onto which you inoculated eight strains of yeast (A-H). The media include a rich medium, an unsupplemented minimal medium, and minimal media each supplemented with one vitamin. Of the yeast strains, one is a prototroph and seven are auxotrophs for a vitamin. After overnight incubation, the following results were observed (tan patches represent growth): D plate 1 (A) B DE F GH plate 5 plate 4 plate 6 Which plate contains an unsupplemented minimal medium? [Select] Which plate contains a rich medium? [Select] plate 2 Which strain is a prototroph? [Select] Strain E is an auxotroph for niacin. Which plate reveals this specific auxotrophy? [ Select] plate 3 plate 7 One strain is an auxotroph for both choline and pantothenic acid. Which one is this most likely to be? [Select]E. coli has a doubling time of 0.345 hours at 37 0C. Starting with 1 milligram of cells, assuming a lag phase of 30 minutes and a stationary phase of 30 minutes, what number of cells would you expect at the end of 13h, assuming that besides the lag and stationary phases the culture is continuously in exponential phase. Assume that ln (2) = 0.69 and that the mass of a single cell is 10^(-12) g.A phenylgalactoside-type compound, which yields galactose when enzymatically cleaved, was added as the sole carbon source to petri dishes containing E. coli culture medium. E. coli can utilize galactose for growth. Each of the following bacterial strains was plated on this medium and scored for growth after 1-2 days. Growth Genotype lacz+ lacI+ lacz+ lacIs lacz+ lacl- + oc lacZ+ lacI+ + a) What do you conclude about this compound with respect to its ability to be cleaved by ß- galactosidase? Why? b) What do you conclude about this compound with respect to its ability to inactivate the Lac repressor? Why? T
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- Sydney Brenner isolated Salmonella typhimurium mutants that were implicated in the biosynthesis of tryptophan and would not grow on minimal medium. When these bacterial mutants were tested on minimal medium to which one of four compounds (indole glycerol phosphate, indole, anthranilic acid, and tryptophan) had been added, the growth responses shown in the following table were obtained. Mutant Minimal medium Anthranilic acid Indole glycerol phosphate Indole Tryptophan trp-1 − − − − + trp-2 − − + + + trp-3 − − − + + trp-4 − − + + + trp-6 − − − − + trp-7 − − − − + trp-8 − + − − + trp-9 − − − − + trp-10 − − − − + trp-11 − − − − + Give the order of indole glycerol phosphate, indole, anthranilic acid, and tryptophan in a biochemical pathway leading to the synthesis of tryptophan. Indicate which step in the pathway is affected by each of the mutations.For the study of alanine production by a recombinant strain of E. coli, cultivation was carried out in a benchtop bioreactor with 4.5 L of culture medium, using glucose as a limiting substrate. During the cultivation, there was no lag phase and the cells showed exponential growth for 5 hours. The following table presents the results of the analysis of ammonia and glucose consumption, and alanine accumulation throughout the cultivation. Knowing that 500 mL of a cell suspension at a concentration of 5.0 g/L (inoculum) was added to the 4.5 L of medium in the reactor and that the YX/NH3 previously determined was 7.5, calculate:a) the maximum specific growth rateb) YX/S and YP/S yield factorsc) How long would it take to reach Cx = 30 g/L if the cells continued with the exponential growth profile until the end of the culture (without nutrient deprivation or any type of inhibition)?d) Describe how the mathematical treatment of the data should be done to determine the type of product formation…Nitrate reduction test can be used to differentiate between Pseudomonads and Enterobacteriaceae. You inoculated your Nitrate broth with your unknown; after incubation, you added 5 drops of reagent A and 5 drops of reagent B. You observe a red color within 2 minutes. Is this a + or a - reaction? Assuming you did not get a red color, you then added a pinch of Zinc and this time tou obtained a red color. What does this mean?