Laboratory 5 Prelab Assignment Name: Affan

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Telek

Laboratory 5.1a*: From Genes to Proteins

1. Using the different DNA Coding Strand sequences provided in Exercise 5.1a, (Table 5.2) transcribe
and translate each; recording your results in Table 5.5a under the correct sequence. Using the codon
table (Table 5.1), determine the primary structure of each peptide. Be sure to always provide your
answer in the 5’ to 3’ direction.

Table 5.5a. Sequences for transcription – translation exercise.

SEQUENCE #1: 5’ TAGAATATGCTTGGTTTCCACAAAAGTTATTACCAAAACTAA 3’
RNA
5 UAG AAUAUGCUUGGUUVC CAC AAAAGUUAVVAC CAAAACUAA3
PROTEIN
Met Leu Gly Phe His Lys Ser Tyr Tyr Gln Asn Stop
SEQUENCE #2: 3’ ATCTTATACGAACCAAAGGTGTTTTCAATAATGGTTTTGATT 5’
RNA
5 UAGAAUAUGCOUGGU UVCCACAAAAGOVAU UACCAAAACUAA3
PROTEIN
Met LeuGly Phe His Lys Ser Tyr Tyr Gln Asn Stop
SEQUENCE #3: 5’ TAGAATATGCTTGGTTTCCACAAAAGTTATTACCAAAACTAA 3’
RNA
5 UAGAAUAUGCOUGGU UVCCACAAAAGOVAUUACCAAAACUAA3
PROTEIN
Met LeuGly Phe His Lys Ser Tyr Tyr Gln Asn Stop
SEQUENCE #4: 3’ ATCTTATACGAACCAAAGGTGTTTTCAATAATGGTTTTGATT 5’
RNA
5 UAGAAUAUGCOUGGU UVCCACAAAAGOVAUUACCAAAACUAA3
PROTEIN
Met LeuGly Phe His Lys Ser Tyr Tyr Gln Asn Stop
Exercise 5.1b*: Mutations

2. Mutations in the DNA sequence in a structural gene can affect the functioning of the protein. Re-
transcribe and translate each of the DNA sequences from Experiment 5.1a with the following
mutations as indicated for each sequence and determine whether the mutation will affect the
functioning of the resulting protein or not.

Sequence #1: Change the eighth A from the left to a G

Sequence #2: Change the second G from the left to a C
Sequence #3: Change the third C from the left to a G
Sequence #4: Change the eleventh A from the left to a C
Sequence 1 Changeof A to G is anexampleofTransitionmutationbecauseAdenineandGuanine are purinesand their s is
interconversion

alledtransitionThisis a silent
m utation
cytosineis pyrmidine Change ofpurineto
pyrimidineiscalledtransue
becauseGuanine isPariseand
equence 2 GtoC is anexample oftransversion 1
equence 3 CtoG is an exampleoftransversionalso
Sequence 4 A is purine and C is pyrimidine
3. Using the following templates, what is the protein for each new sequence?

Table 5.5b. Sequences for translation to protein exercise.

New Sequence # 15 TAGAATATGCTTGGTTTCCACAAGAGTTATTACCAAAACTA.AZ
New RNA
5 UAGAAUAVGCUU GGUUVCCACAAGAGUUAVVACCAAAACUAA 3
Protein
Met Leu Gly Phe His Lys Ser Tyr Tyr Gin Asn Stop
New Sequence # 2 3
ATCTTATACGAACCAAACGTGTTTTCAATAATGGTTTTGATT 5
New RNA
5 UAGAAUAUGCUUGGU UUG CACAAAAGUVAUVACCAAAACUAA 3
Protein
Met Leu Gly Leu His Lys Ser Tyr Tyr GlnAsn Stop
New Sequence # 3 S TAGAAT
ATGCTTGGT TTC GACAAA AGTTATTACCAAAACTAA3
New RNA
5 UAGAAUAUGCUUGGU UUC GACAAA AGUUAVVACCAAAACUAA 3
Protein
Met Leu Gly Phe Asp Lys Ser Tyr Tyr GinAsn Stop
New Sequence # 4 3
ATCTTATACGAACCAAAGGTGTTTTCAATCATGGTTTTGATT S
New RNA
5 UAGAAUAUGCUU GGU UVCCACAAAAGUUAGUACCAAAACUAA 3
Protein
Met Leu Gly Phe His Lys Ser stop
4. What kind of a mutation is each one an example of?
/Sequence #1 silent mutation

Sequence #2 Missensemutation

Sequence #3 Missense mutation

Sequence #4Nonsense mutation

5. Predict whether each of these changes will affect the function of the protein. Why or why not?

Sequence #1 No as theSame aminoacid sequence willbecoded

Sequence #2Yes there is achange CACMRNAsequence isCTG t Gta mRNA sequence is Gtc

Sequence #3 Yesthere is achangeAAGmRNAsequence isTTC GAGmRNAsequence is CTC

Sequence #4Yesthere TCAmRNAsequence is AGT

is achangeACAmRNAsequence isTGT

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Laboratory 5.2*: Practice with Punnett Squares

Exercise 5.2: Using Punnett Squares

Imagine a man has freckles and a woman (his partner) does not have freckles. Freckles are a dominant
trait. We will denote the allele for freckles as Q and the allele for no freckles as q. The man’s mother did
not have freckles, and the woman’s father did have freckles.

6. Record the genotype for each individual:

The woman
The man qq
Qq
7. Based on the genotype for each individual, give the possible gametes each could produce

The woman and

The man q and q
Q q
8. Imagine this couple has a child. Complete the Punnett Square to determine the probability the child,
born to this couple will have freckles. 50 Chancethatthiscouple'schildwillhavefreckles 214 41 507
woman
man q q
Qq Qq
I qq qq
9. What is the probability that the child, born to this couple, will be homozygous dominant?
0 as forthistoevenbepossible boththemotherandfatherwouldhavetobe atleastheterozygousdominant
10. Provide the genotype ratio:
Qaqq 2 2

and phenotype ratio:Freckles NoFreckles 2 2

Imagine another couple similar to above, but this time we will also consider hair. This man has freckles
and straight hair. The woman has no freckles and has curly hair. Freckles (Q) are dominant to no freckles
(q). Curly hair (H) is dominant to straight hair (h).

The woman has the following genotype: qqHh

The man has the following genotype: Qqhh

11. Based on the genotype for each individual, give the possible gametes each could produce

The woman
The man
att and
and qt and
and
ah and
and
ah
ah ah ah ah

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12. Imagine this couple has a child. Complete the Punnett Square to determine the probability that the
child born to this couple will have freckles and curly hair. What is that probability?
4112 44 254

woman
man
ah ah ah ah
Oh Qalth Qalth Qqhh Qahh
QL Qalth Qalth Qghh Qqhh
gh walth qglth qqhh ahh
qh 99th auth qqhh qqhh
Genotype ratio:
44 14
Phenotype ratio: 251FrecklesCurlyHair 25 Freckles Straight Hair 25
NoFrecklesCurlyHair
251 No Freckles StraightHair
13. You observe several different ladybugs in your garden and decide to cross purple winged ladybugs
with yellow dots to see what offspring will be produced. Fill in the Punnet square assuming that the
ladybugs chosen are heterozygous in both traits. Use the letters B and b to describe the alleles for wing
color, whereas B represents purple wings that are dominant to b (red wings). Use D and d to describe
the alleles for dot color where D represents the dominant yellow dots and d represents the black
recessive dots. Remember the capital letter signifies the dominant allele and the small letter signifies
the recessive allele.

What phenotypes will be present? (Notice what the question is asking and answer appropriately)

BD Bd BD bd
BD BBDD BBDd BDD BbDd
Bd BBDd BBdd BbDd Bbdd
bD BbDD BbDd bbDD bbDd
bd Bbbd Bbdd bbDd bbdd
The possible phenotypes that willbepresent intheoffspring are Purple Wing YellowDots
PurpleWingBlackDotsPurpleWings BlackDots RedWings YellowDots andRed

Wings Black Dots

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Experiment 5.3: Examining Prepared Slides of Plant and Animal cells undergoing Mitosis

14. Using the space provided below, make drawings of each phase of mitosis for both plant and animal
cells in Figure 5.1 a to j making note of the questions asked in manual. These drawing are part of your
of Prelab assignment and are to be done before coming to the lab so that you are familiar with the
various stages of mitosis and know what you should be looking for when you come to use the
microscope in the lab. Make these drawing in PENCIL so that changes can be made to them as to what
you actually see under the microscope. Be prepared to identify these phases in the future, so make
accurate detailed drawings of what you see. Again, once you look under the microscope, you can make
adjustments to these if needed. It will be what you see in the microscope, not the textbook that you
will be tested on.

Exercise 5.3a: Mitosis in Plant Cells Exercise 5.3b: Mitosis Animal Cells

Nucleus
ohhh
ooooo
Nucleus

i
Figure 5.1a. Plant cell in Interphase Figure 5.1b. Animal cell in Interphase
Magnification: 440 Magnification: 440

Nucleusdisspeaing
d Chromosomes
dense

im Ifqf
yychrgqgtn
yyyy.gg

Wow it j i naaman
dissapates

Figure 5.1c. Plant cell in Prophase Figure 5.1d. Animal cell in Prophase
Magnification: 440 Magnification: NO

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 Fullycoinedchromosomes
arelinedup inthe

am
IfYÉtgygf.de
ffgg ostgiissi
Figure 5.1e. Plant cell in Metaphase Figure 5.1f. Animal cell in Metaphase
Magnification: 440 Magnification: NO
chromosome
chromosome dividedinto two
chromosomes
Yeperated
In Its www
i
amassing
tEEEE
Figure 5.1g. Plant cell in Anaphase Figure 5.1h. Animal cell in Anaphase
Magnification: NO Magnification: 440
chromosome
reachedpoleand begin

DayIting
qq
if
division into
twoiantia
daughtercens

Figure 5.1i. Plant cell in Telophase Figure 5.1j. Animal cell in Telophase
Magnification: Magnification:
NO NO

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15. What is the distinguishing feature of each of the stages of mitosis, including interphase, in plant cells
(as represented by Allium) and animal cells (represented by Whitefish blastula)? Use Table 5.6 to
answer this question. Do not simply list the characteristics given in the manual, but rather makes notes
that will be useful to you when using this material to study from.

Table 5.6: Distinguishing characteristics of different stages of cell cycle

Phase Distinguishing feature(s) in Allium Distinguishing feature(s) in White Fish

(plant cells)
- cell grows and makes a copy of its
DNA
- chromosomes uncoil into long, thin structures
and no longer appear as individual threads
Interphase - cell is either growing and preparing to divide
or doing a specialized function in a multicellular
species
- Golgi apparatus acquires enzymes, structural
proteins, and glucose molecules
Blastula (animal cells)
- The three stages of interphase are called G1, S,
and G2.
- In the G1 phase, the cell is getting the building
blocks of DNA and the related proteins
- also getting a su cient amount of energy to
complete the task of replicating each chromosome
in the nucleus
- S phase (synthesis phase), the formation of sister
chromatids that are rmly attached at the
centromere region.
- each chromosome is made of two sister
chromatids and is a duplicated chromosome

- chromosomes continue to condense

- chromosomes condense and - Centromeres move toward the
become visible opposite poles
Prophase - spindle bers emerge from the
centrosomes
- nuclear envelope breaks down
- nucleolus begins to disappear

- spindle is fully developed - sister chromatid attaches to a

- centromeres are on opposites poles spindle ber that initially came from
opposite poles
Metaphase

- sister chromatids (chromosomes) - non-kinetochore spindle bers grow

are pulled apart to opposite poles and make the cell longer
- cohesin proteins that had binded the
Anaphase sister chromatids break down

Cytokinesis: a cell plate separates the daughter cells Cytokinesis: a cleavage furrow separates the daughter
- Golgi vesicles move on microtubules to collect at the cells
metaphase plate - chromosomes are on opposite ends of poles and
- cell plate: vesicles fuse from the center toward the cell decondense
Telophase walls
- more vesicles fuse = cell plate
- nuclear envelope surround chromosomes
- mitotic spindle breaks down
getting bigger until it merges with the cell wall
- enzymes use glucose (obtained between the membrane
lavers) to build a new cell wall of cellulose
- Golgi membranes become the plasma membrane on
either side of the new cell wall