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: 14333

#Section 5: Bioprocess Engineering and Process Biotechnology

A cylindrical bioreactor of diameter 3 m has four baffles. A Rushton turbine mounted in the reactor has a diameter one third the tank diameter and is operated at a speed of 90 rpm. The density of the fluid is approximately I g cm^-3. The reactor is used to culture an anaerobic organism that does not require gas sparging. The broth can be assumed Newtonian. As the cells grow, the viscosity of the broth increases. when the viscosity is 1000 times greater than water, estimate the power required to achieve turbulence.

#Question id: 14334

#Section 5: Bioprocess Engineering and Process Biotechnology

Laboratory-scale fermenters are usually mixed using small stirrers with electric motors rated between 100 and 500 W. One such motor is used to drive a 7cm turbine impeller in a small reactor containing fluid with the properties of water. The stirrer speed is 900 rpm. Estimate the power requirements for this process.

#Question id: 14336

#Section 5: Bioprocess Engineering and Process Biotechnology

A cylindrical stirred bioreactor of diameter and height 2 m has a Rushton turbine one-third the tank diameter in size. The bioreactor contains Newtonian culture broth with the same density as water and with viscosity 4 cP. The tank is now aerated. In the presence of gas bubbles, the approximate relationship between ungassed power number (N_P)₀ and gassed power number (N_P)g is: (N_P)g =0.5 (N_P)₀. What maximum stirrer speed is now possible in the sparged reactor? Estimate the mixing time.

#Question id: 14337

#Section 5: Bioprocess Engineering and Process Biotechnology

#Question id: 14338

#Section 5: Bioprocess Engineering and Process Biotechnology

Quasi steady state condition is