19) Core Concept: Information Flow- Explain how information from DNA results in the synthesis of a protein. Explain how information in the environment ultimately results in changes to gene expression.

20) Explain the steps in DNA recombinant technology (FROM GFP LAB). Include restriction enzymes, plasmids, heat shock, transformation and names of any enzyme that is needed in your answer.

21) Explain the purpose and/or what is happening at any step in the GFP lab protocols.

22) Predict how defects in the regulation of cell-cycle checkpoints would affect cells and explain how these defects relate to uncontrolled division in cancer and cancer progression.

The lacI gene regulates transcription of the structural genes by producing a repressor molecule that is allosteric, meaning that

1. Which of the following are TRUE about sperm?

Select one or more:

a. Hyaluronidase is the enzyme involved in dissolving the protective coating around the unfertilized ovum.

b. The first sperm cell to reach the egg successfully fertilizes that cell.

c. Sperm cells have half the number of chromosomes as a spermatagoium and are about the same size.

d. Sperm cell capacitation, which occurs after ejaculation, is required for sperm cells to be active.

e. Mitochodria are located in the mid piece of the tail below the head.

f. The head of the sperm contains the DNA from the father.

g. The acrosome is located at the base of the sperm cell's tail.

16. Why does FADH2 provide energy for 1.5 ATP molecules in the ETC, whereas NADH provides energy for 2.5 ATP molecules? ( Show calculations. )






17. How does each of the following regulate the CAC:

       a/ high levels of NADH
     
      
       b/ high levels of ATP
     
      
       c/ high levels of ADP

      
       d/ low levels of NADH




18. Where is ATP synthase for oxidative phosphorylation located in the cell?




19. Why do H+ leave the intermembrane space and return to the matrix of the mitochondrion?





20. Why do the enzyme complexes that pump H+ extend across the mitochondrial membrane from the matrix to the intermembrane space?

The reaction A + B → C has a ∆G^0’ of -20 kJ/mol at 25 ⁰C. Starting under

       Standard Conditions, one can predict that:

A. at equilibrium, the concentration of B will exceed the concentration of A.

B. at equilibrium, the concentration of C will be less than the concentration of A.

C. at equilibrium, the concentration of C will be much greater than the

     concentration of A or B.

D. C will rapidly break down to A + B.

E. when A and B are mixed, the reaction will proceed rapidly toward formation

     of C.

29. Estimate the value of K_m for an enzyme-catalyzed reaction for which the

      following data points were obtained.

[S] (M)                       V₀ (mM/min)

2.5 x 10^-6                                28

4.0 x 10^-6                                40

1.0 x 10^-5                                70

2.0 x 10^-5                                95

4.0 x 10^-5                                112

1.0 x 10^-4                                128

2.0 x 10^-3                                139

1.0 x 10^-2                                140

A. 1.0 x 10^-4

B. 1.0 x 10^-3

C. 1.0 x 10^-6

D. 1.0 x 10^-5

E. 1.0 x 10^-7

30. pH = pK_a when:

[A^-]/[HA] = 0

log ([A^-]/[HA]) = 1

[A^-] >> [HA]

[A^-] = [HA]

log ([HA]/[A^-]) = 1

31. In the conversion of A into D in the following biochemical pathway, enzymes

       E_A, E_B, and E_C have the K_m values indicated under each enzyme. If all of the

      substrates and products are present at a concentration of 10^-4 M and the

      enzymes have approximately the same V_max which step will be rate limiting?

                                         R_x1      R_x2     R_x3

                                                  A   ↔   B   ↔   C ↔ D

                                                          E_A        E_B       E_C

                                                           K_m =     10^-2M    10^-4M   10^-4M

A. Rx 3         

B. Rx 1         

C. Rx 2         

D. Rx’s 1 and 3

E. Rx’s 2 and 3       

32. Consider the following K_eq values with the appropriate ΔG^0’?

                                       K_eq                                        ΔG^0’

                        A           1.0                           1      28.53

                      B           10^-5                             2     -11.42

                        C           10⁴                          3      5.69

                        D           10²                          4       0

                        E           10^-1                         5      -22.84    

A K_eq of 10² would correspond to which of the following ΔG^0’ values?

28.53

-11.42

5.69

0

-22.84

33. For an enzyme that follows simple Michaelis-Menten kinetics, what is the

      value of V_max if V₀ is equal to 1 µmol minute^-1 when [S] = 1/10 K_m?

11.0 µmol minute^-1

1.10 µmol minute^-1

0.10 µmol minute^-1

10.1 µmol minute^-1

1.0   µmol minute^-1

Purpose

        The clear understanding of energy transformations (Bioenergetics and Photosynthesis) by cells explains how cells can maintain their homeostasis in a continuously changing world. Completing this assignment will assist you in developing clarity of how cells function energetically on the planet and how some cells capture sunlight to build organic molecules. These principles of Bioenergetics and Photosynthesis will help clarify the Course Content Learning Outcomes number 2 (Understand the basic principles and ideas of chemistry and organic chemistry) and 4 (Able to explain the concept of a cell and describe the functions of its parts) for this course. This exercise will also clarify the Campus Wide Outcome of Critical Thinking (See page 3 (Critical Thinking) of the syllabus). The knowledge gained could assist students in other related science courses and courses in Psychology, Criminal Justice, and others.

Task

        Armed with a textbook (Chapters 5 & 6), the Bioenergetics and Photosynthesis Power Point presentations, or other reliable sources answer the following questions about Bioenergetics and Photosynthesis. Be creative in how you approach and answer each question. Do not just copy and paste information from another source. Read the source and then write your answer in your own words so that it becomes your answer. Remember in science there are no “correct” answers but rather explanations of phenomena. This assignment will be due .....

Criteria

        This assignment is worth .... total. The possible points for each question are in parenthesis after the question number. A complete answer to each question will involve being cognizant of the principles of Bioenergetics (The energy flow through a biological system.) and Photosynthesis (The harvesting of sunlight energy to build organic molecules.). Clear answers will have the involved cellular parts and molecules labeled clearly with descriptions and definitions that are clear, concise, and will contain enough detail that anyone reading your answer could potentially draw the same conclusions that you have by answering the questions.

1. (5) Define the following terms as they relate to an enzyme: activation energy, active site,

conformation, denaturation, and coenzyme/cofactor binding site.

2. (5) How are enzymes inhibited by competing and by non-competing molecules?

3. (5) Draw a graph that would represent enzyme function at various temperatures and draw a

graph that would represent enzyme function at various pH levels.

4. (5) Describe and give an example of the concept of energy coupling reactions in regards to cellular

ATP usage.

5. (10) Describe the light dependent reactions of photosynthesis in five steps or less.

6. (10) Describe the light independent reactions (Calvin cycle) of photosynthesis in five steps or less.

Divide the cycle into two halves: Molecule Building and Rearrangement before listing your steps to help focus your thoughts.

Q1 A research laboratory identified a gene X of medicinal value in a plant species. You are given a small fragment of DNA containing gene X and the cloning vector pZoom. Maps of the 4 kb cloning vector pZoom and the 10 kb plant DNA fragment are shown in Figure 1 and 2, respectively. The PCR primer pairs (shown in Figure 1) F1/R1, F2/R2 and F3/R3 amplify fragments of 0.1, 0.6 and 0.8 kb, respectively. The antibiotic resistance gene A and gene B code for chloramphenicol and streptomycin resistance, respectively. The three restriction enzymes that cut this vector are BamHI, XbaI and HindIII represented on the map as enzymes I, II and III, respectively. You are given purified DNA of both the vector and the gene X containing DNA fragment at 0.2 µg/ul. Create a recombinant plasmid containing the complete gene X in the provided cloning vector pZoom. a. Design restriction digestion reactions using appropriate enzymes in such a way that you get a final concentration of 50 ng/µl for the digested vector and for the plant DNA (insert DNA) in the reaction. Your digestion should include all components in a 20 µL reaction. All enzymes are supplied with a concentration of 10 units/µL; you may use 1 µL of the enzyme in each reaction. Buffers for each enzyme are available as 10 times concentrated (10X) stocks. b. Generate three ligation reactions with 1:1, 2:1 and 3:1 molar ratios of the insert and the vector DNA. Your ligation should include all components in a 30 µL reaction. Keep the vector amount fixed at 100 ng per ligation reaction. You are provided with 10 x ligase buffer and DNA ligase (0.5 U/µL) to set up your ligations. c. Develop a strategy to select transformants and a quick method to screen recombinants; show your screening method using a figure. Predict the expected results with an explanation. d. Devise a restriction analysis method to confirm the desired recombinant; use a single most appropriate enzyme. Calculate the expected sizes of the restriction fragments from each, the vector and the desired recombinant. e. Devise a PCR strategy to confirm the desired recombinant. Calculate the expected sizes of the PCR fragments from the desired recombinant in a multiplex PCR including all three primer pairs. How will these sizes differ from the same multiplex PCR if the template is vector DNA instead of the recombinant plasmid?

Economists refer to the talents, training, and education of workers as

Multiple Choice

• human capital.

• physical capital.

• average labor productivity.

• labor supply.