Download pGlo Lab write-up materials

Diff Biology: pGlo Bacterial Transformation Lab
Names:
I. pGlo Bacterial Transformation Pre-lab Questions.
1. What is the purpose of this lab? What are we trying to do?
2. What is the original source of green fluorescence protein?
a. Ultra-violet Light
b. Humans
c. Jellyfish
d. Fireflies
3. Complete the following sentences using the terms listed:
Bacterial Transformation
LB Nutrient Broth
Plasmid
Genetic Regulation
Calcium Chloride (CaCl2)
Genetic Selection
Heat Shock Treatment
a. A ____________________________ is a small, circular piece of DNA that can multiply on its own.
b. ___________________________ is a sterile solution that makes cells able to take in a plasmid (competent).
c. Bacteria can be grown under different conditions to determine what traits they have by using ___________________.
d. ________________________________ is the process of moving genes from one organism to another.
e. _________________________________ is a mixture that allows cells to grow and produce gene products (proteins).
f.
Competent cells take in plasmids during _______________________________ in a transformation procedure.
g. Organisms can control the kinds and amounts of proteins they produce because of the process of ________________.
6. Normal, non-transformed, E.coli bacteria are white, use arabinose (Ara) as a food source, and cannot live when
exposed to antibiotics like ampicillin (amp). What will you see if E.coli is grown under the following
conditions? Will there be growth or no growth? Circle your answer below for each plate.
LB Nutrient Agar +
Ampicillin:
LB Nutrient Agar + Ampicillin
+ Arabinose:
growth
growth
LB Nutrient Agar:
growth
no growth
no growth
no growth
7. What do you expect to see if E.coli bacteria is transformed so that it produces a green fluorescence protein in
the presence of arabinose and the E.coli is now resistant to ampicillin? Will there be growth or no growth?
Will there be any glowing bacteria? Circle your answer below for each plate.
LB Nutrient Agar + Ampicillin:
growth
no growth
glowing
not glowing
LB Nutrient Agar + Ampicillin +
Arabinose:
LB Nutrient Agar:
growth
no growth
growth
no growth
glowing
not glowing
glowing
not glowing
II. pGlo Bacterial Transformation: Experimental Observations
1. Review your predictions from the Pre-Lab worksheet for bacterial growth. What are your actual results? Under
each plate below, list your observations. (If you did not end up with any glowing bacteria, indicate this in your
observations, by placing an “X” over the plates below. Fill in the observations by using another group’s results.)
Pattern of growth:
(no growth,
single colonies,
or lawn)
Average size of
colonies (mm):
Color of colonies
under normal
lighting:
Color of colonies
under UV light:
Number of
colonies:
Additional Notes:
2. Which traits that you initially observed for E.coli did not change?
3. Which traits that you initially observed for E.coli did change after performing the procedure?