To express a 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 do the digestion of the insert and the vector and then ligate the two digestions together. You then transform the ligation into bacteria and select for ampicillin resistance. You get three colonies on your transformation plate. You isolate plasmid from each one and cut each plasmid with the enzyme XbaI. You then run your three digestions on an agarose gel and see the following patterns of bands. Describe what each plasmid actually was that was contained in each of the three colonies.

 

What is the Colony 1’s plasmid is;

#Question id: 14313

#Section 5: Bioprocess Engineering and Process Biotechnology

An industrial waste-water stream is fed to a stirred-tank reactor continuously and the cells are recycled back to the reactor from the bottom of the sedimentation tank placed after the reactor. The following are given for the system: F = 100 l/h; S0 = 5000 mg/l; mm = 0.25 h-1 ; Ks = 200 mg/l; α (recycle ratio) = 0.6; C (cell concentration factor) = 2; Y ^M _X/S = 0.4. The effluent concentration is desired to be 100 mg/l.  Determine the cell concentration in the reactor and in the recycle stream.

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#Question id: 14314

#Section 5: Bioprocess Engineering and Process Biotechnology

An industrial waste-water stream is fed to a stirred-tank reactor continuously and the cells are recycled back to the reactor from the bottom of the sedimentation tank placed after the reactor. The following are given for the system: F = 100 l/h; S0 = 5000 mg/l; mm = 0.25 h^-1 ; Ks = 200 mg/l; α (recycle ratio) = 0.6; C (cell concentration factor) = 2; Y ^M _X/S = 0.4. The effluent concentration is desired to be 100 mg/l.  The effluent concentration is desired to be 100 mg/l. If the residence time is 2 h in the sedimentation tank, determine the volume of the sedimentation tank and cell concentration in the effluent of the sedimentation tank.

#Question id: 14315

#Section 5: Bioprocess Engineering and Process Biotechnology

Anaerobic digestion of volatile acids by methane bacteria is represented by the equation:

The composition of methane bacteria is approximated by the empirical formula CH_1.4O_o.40N_0.20. For each kg acetic acid consumed, 0.67 kg CO ₂ is evolved. How does the yield of methane under these conditions compare with the maximum possible yield?

#Question id: 14316

#Section 5: Bioprocess Engineering and Process Biotechnology

Cellulomonas bacteria used as single-cell protein for human or animal food are produced from glucose under anaerobic conditions. All carbon in the substrate is converted into biomass; ammonia is used as nitrogen source. The molecular formula for the biomass is CH_I.560_0.54N_0.16; the cells also contain 5% ash. How does the yield of biomass from substrate in mass and molar terms compare with the maximum possible biomass yield?

#Question id: 14317

#Section 5: Bioprocess Engineering and Process Biotechnology

system for manufacture of single-cell protein is Methylophilus methylotrophus. This organism is produced aerobically from methanol with ammonia as nitrogen source. The molecular formula for the biomass is CH _1.680_0.36N_0.22; these cells contain 6% ash. What is the maximum possible biomass yield from methanol?

#Question id: 14318

#Section 5: Bioprocess Engineering and Process Biotechnology

system for manufacture of single-cell protein is Methylophilus methylotrophus. This organism is produced aerobically from methanol with ammonia as nitrogen source. The molecular formula for the biomass is CH _1.680_0.36N_0.22; these cells contain 6% ash. If the actual yield of biomass from methanol is 42% the thermodynamic maximum, what is the oxygen demand?