You are studying a specific gene in yeast, and you want to express that yeast gene in E. coli. Your task is to design a strategy to insert the yeast gene into the bacterial plasmid. Below is a map of the area of the yeast genome surrounding the gene in which you are interested.

 

The distance between each tick mark placed on the line above is 100 bases in length

Below are the enzymes you can use, with their specific cut sites shown 5’-XXXXXX-3’ 3’-XXXXXX-5’

 

The plasmid is 5,000 bases long and the two farthest restriction enzyme sites are 200 bases apart. The plasmid has an ampicillin resistance gene somewhere on the plasmid distal from the restriction cut sites.

                                    

You transform your ligation planned in which two restriction enzymes would you use to design a way to get the insert into the vector if you had to use two enzymes simultaneously, into bacteria and plate the bacteria on Petri plates containing ampicillin. (You actually transform six different ligation mixtures, which are described below, into six different populations of cells, and plate each transformation onto a different plate, because you want to do all of the correct controls.) The next day you come in to lab to look at how many colonies of bacteria are on each plate. You are really excited, because the number of colonies you see on each plate tells you that the entire procedure worked! Which of the three following patterns of number of colonies did you see in order to conclude that you had a successful transformation?

In this table, DV = digested vector. DYG = digested yeast genome.

 

#Question id: 14322

#Section 5: Bioprocess Engineering and Process Biotechnology

Production of recombinant protein by a geneticallyengineered strain of Escherichia coli is proportional to cell growth. Ammonia is used as nitrogen source for aerobic respiration of glucose. The recombinant protein has an overall formula CH_I.550_0.31N_0.25. The yield ofbiomass from glucose is measured at 0.48 g g^-1; the yield of recombinant protein from glucose is about 20% that for cells. How much ammonia is required per gmol glucose?

#Question id: 14323

#Section 5: Bioprocess Engineering and Process Biotechnology

Production of recombinant protein by a geneticallyengineered strain of Escherichia coli is proportional to cell growth. Ammonia is used as nitrogen source for aerobic respiration of glucose. The recombinant protein has an overall formula CHI.5500.31N0.25. The yield ofbiomass from glucose is measured at 0.48 g g-1; the yield of recombinant protein from glucose is about 20% that for cells. What is the oxygen demand per gmol glucose?

#Question id: 14324

#Section 5: Bioprocess Engineering and Process Biotechnology

Production of recombinant protein by a geneticallyengineered strain of Escherichia coli is proportional to cell growth. Ammonia is used as nitrogen source for aerobic respiration of glucose. The recombinant protein has an overall formula CHI.5500.31N0.25. The yield of biomass from glucose is measured at 0.48 g g^-1; the yield of recombinant protein from glucose is about 20% that for cells. If the biomass yield remains at 0.48 g g^{- l}, how much change in oxygen requirement for wild-type E. coli unable to synthesise recombinant protein?

#Question id: 14325

#Section 5: Bioprocess Engineering and Process Biotechnology

Production of recombinant protein by a geneticallyengineered strain of Escherichia coli is proportional to cell growth. Ammonia is used as nitrogen source for aerobic respiration of glucose. The recombinant protein has an overall formula CHI.5500.31N0.25. The yield ofbiomass from glucose is measured at 0.48 g g-1; the yield of recombinant protein from glucose is about 20% that for cells. If the biomass yield remains at 0.48 g g- l, how much change in ammonia requirement for wild-type E. coliunable to synthesise recombinant protein?

#Question id: 14326

#Section 5: Bioprocess Engineering and Process Biotechnology

The chemical reaction equation for conversion of ethanol (C₂H₆0) to acetic acid (C₂H₄0₂) is:

Acetic acid is produced from ethanol during growth of Acetobacter aceti, which has the composition CH_1.80_0.5N_0.2. Biomass yield from substrate is 0.14 g g^{- 1}; product yield from substrate is 0.92 g g^-1. Ammonia is used as nitrogen source. How much gmol of oxygen is demanded for acetic acid production?

#Question id: 14327

#Section 5: Bioprocess Engineering and Process Biotechnology

Bakers' yeast is produced in a 50 000-litre fermenter under aerobic conditions. The carbon substrate is sucrose; ammonia is provided as nitrogen source. The average biomass composition is CH_1.830_0.55N_0.17 with 5% ash. Under efficient growth conditions, biomass is the only major product; the biomass yield from sucrose is 0.5 g g^-I. If the specific growth-rate is 0.45 h^-l, estimate the rate of heat removal required to maintain constant temperature in the fermenter when the yeast concentration is 10 g l^{- 1}     _______________________.10^6