6c) Explain how eukaryotic organelles are able to transport materials between one another.

6d) Considering your answer from 6c, explain how cells are able to import and export large and larger amount of molecules across the plasma membrane.

You have isolated a strain of brewer’s yeast (Saccharomyces cerevisiae) in the lab for your second job at a hip new microbrew . You find that this strain ferments more efficiently and adds a superior flavor profile to your brews. You want to clone the strain of yeast and generate a genomic library to determine the genes responsible for this finding. To do so you:

1. Obtain fragments of the whole yeast genome (DNA) through restriction digestion
2. Open your vector and ligate the DNA fragments
3. Transform E. coli with these vectors
4. Select for E. coli that have obtained vectors
5. Screen for E. coli transformed with recombinant plasmids
6. Obtain recombinant plasmids from the library and transform yeast with them
7. Clone by complementation the gene that can give superior fermentation.

Note: the cloning site is within the lacZ gene.

a) (3 points) In step 3 you transform E.coli with your plasmids and plate them on a solid agar medium to grow and isolate colonies. What medium should you used to distinguish the bacteria transformed with recombinant vectors from ones that carry non-recombinant vectors? Briefly explain how the medium allows for you to distinguish. (hint: look up the substrate X-gal)

b) (2 points) You identify a recombinant vector that passes your complementation test. You are curious whether this vector will cause bacteria to ferment sugars for brewing. Provide and explain two reasons why this recombinant vector will NOT work in bacteria.

. For each of the restriction enzymes listed below: (i) Approximately how many restriction fragments would result from digestion of the human genome (3 x 109 bases) with the enzyme? (ii) State whether the fragments produced by digestion with each enzyme would have sticky ends with a 5’ overhang, sticky ends with a 3’ overhang, or blunt ends. (The recognition sequence for each enzyme is given in parentheses, where N means any of the four nucleotides. ^ marks the site of cleavage.)

a. EcoRV (GAT^ATC)

b. HpaII (C^CGG)

c. DrdI (GACNNNN^NNGTC)

(h) Pseudomonas aeruginosa also uses the Entner-Doudoroff pathway to degrade glucose. Use IMG or BLAST (or MetaCyc) to identify which enzymes in the EMP pathway are missing from P. aeruginosa PAO1.

(i) Biochemical experiments performed by Robertson and McCullough (1968) suggested that Brucella abortus uses the pentose phosphate pathway, instead of the EMP or Entner-Doudoroff pathway, to degrade glucose. The labeling and enzymological results, obtained with B. abortus strain 19 grown in rich media in shaken culture, suggested that key enzymes are missing. Use IMG or BLAST (or MetaCyc) to identify which enzymes in the EMP and ED pathways are missing from B. abortus S19. Is there a discrepancy from experimental results? Explain.

Please only answer if you know the answer. Appreciated

In what polysaccharide does a redwood tree store its energy?

Describe the experimental procedure used to accomplish DNA transformation into bacterial cells.

Briefly describe the 3 signals involved in the activation and differentiation of a naive CD4+ T cell into an effector Th2 (T helper) cell.

Identify which Mill's method is best exemplified. Explain why the Mill's method is exemplified. Demonstrate your understanding of the scientific concept and the logical structure by writing for each passage why the particular Mill's method is best exemplified.There may be more than one method in each passage.

1. The Sac locus was recently shown by linkage analysis and genetic rescue to encode T1R3, thus implicating a member of the T1r gene family in sweet taste detection. Indeed, functional expression studies in heterologous cells revealed that T1R3 combines with T1R2 (T1R2+3) to form a sweet taste receptor that responds to all classes of sweet tastants, including natural sugars, artificial sweeteners, d-amino acids and intensely sweet proteins.

2.Humans and mice show some prominent differences in their ability to taste certain artificial sweeteners and intensely sweet proteins — for example, mice cannot taste aspartame or monellin. Notably, introduction of the human T1R2 receptor into mice significantly changes their sweet taste preferences to a human-like response profile, proving that species differences in sweet taste sensitivity and selectivity are a direct reflection of T1R-sequence variation between species.

1. What is the difference between transcription and DNA replication?
2. In what phase of the cell cycle does DNA replication occur?
3. What is the difference between Mitosis and cytokenesis?
4. Give the function of the following:
   • DNA polymetase
   • Helicase
   • binding proteins
   • ligase
   • primase
5. What is a sister chromatid?
6. What is the function of a centrosome?

The phenotypic ratio of the F₁ offspring produced by the cross between the tan body female fly and the wild-type male fly is 50% wild females: 50% tan body males.What chromosome did the F₂ offspring receive from their mother (cross between the tan body female fly and the wild-type male fly)?

Select one:

a. Y chromosome with no allele for body color

b. X chromosome with a wild allele for body color

c. X chromosome with either a wild or a tan allele for body color

d. X chromosome with a tan allele for body color

1. In a Quelling reaction test, the antibodies added to the sputum sample are reacting to what morphological feature on the bacterium which causes the bacterium to swell - a positive test result.

outer membrane

flagellum

capsule

cell wall

2. When the Western Blot is used to test for Lyme Disease what the procedure is testing for specifically in the individual is the presence of

  fragments of the bacterial outer membrane

the bacterium

specific DNA sequences from the bacterium

antibodies against the bacterium

3. Restriction enzymes, used in DNA fingerprinting, search for and cut pieces of ds DNA that are palindromes. Which below is a palindrome? (In the answers the top strand is listed first followed by it's complement)

CCCCCC GGGGGG

AAATTG TTTAAC

GAATTC CTTAAG

GTAAGC CATTCG

For which of the following medical conditions is the interaction of genes and environment likely to be important in the manifestation of the phenotype? A) alcoholism B) type 2 diabetes C) obesity D) A and C E) All of the above