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.

 

Which colony’s plasmid do you actually want to use for your studies?

#Question id: 536

#Unit 1. Molecules and their Interaction Relevant to Biology

Glucose 6-phosphate allosterically inhibits

A) hexokinase I.

B) glucokinase.

C) hexokinase II.

#Question id: 537

#Unit 1. Molecules and their Interaction Relevant to Biology

Substrate level phosphorylation

A) describes the conversion of ADP into ATP with the addition of inorganic phosphate every place throughout the cell.

B) describes the formation of ADP by phosphoryl group transfer from 1,3 bisphosphoglycerate.

C) is the formation of ATP by phosphoryl group transfer from a higher energy compound.

#Question id: 538

#Unit 1. Molecules and their Interaction Relevant to Biology

Which of the following statements are correct about cells that form ATP mainly by glycolysis are

A) anaerobic yeasts.

B) lactic acid bacteria.

C) kidney medulla cells.

#Question id: 539

#Unit 1. Molecules and their Interaction Relevant to Biology

Which of the following mutases catalyze the formation of a 2,3-BPG intermediate?

A) muscle phosphoglycerate mutases

B) plant phosphoglycerate mutases

C) yeast phosphoglycerate mutases

#Question id: 540

#Unit 1. Molecules and their Interaction Relevant to Biology

During strenuous exercise, the NADH formed in the glyceraldehyde 3-phosphate dehydrogenase reaction in skeletal muscle must be reoxidized to NAD+ if glycolysis is to continue.  The most important reaction involved in the reoxidation of NADH is:

#Question id: 541

#Unit 1. Molecules and their Interaction Relevant to Biology

The overall △G for glycolysis is -72 kJ/mol in erythrocytes. Which statement below is true?

A) The value of △G0ʹ is also -72 kJ/mol since the cytosol pH is close to 7.

B) The free energy of glycolysis is found as the sum of the standard free energy changes for

the individual pathway reactions.

C) The negative sign of △G shows that this pathway will proceed toward product (pyruvate) under normal cellular conditions.