A Technical Report About
Mutation
Perez, Ma. Betina Andrea P.
Q3A1
Genetics Laboratory
Ms. Tin Manansala
May 24, 2017
Introduction:
Mutations are the alteration of the genetic sequence, which is the root why organisms
are different from each other. Mutation affects the organisms descendants because it occurs in
cells that produce the next generation and affect the hereditary material. In addition, mutation is
a change that influences the nucleic acids. Nucleic acids are the building block of the DNA
which contains our genetic material (Loewe, 2008). Mutations are heritable and permanent
(Ramirez et al, 2013)
Chromosome mutations happen for two reasons, numerical and by its genome structure.
There are two types of chromosomal mutation according to its number, aneuploidy and
euploidy. In euploidy, the chromosome changes the number of it in whole sets. While in
aneuploidy, mutations happen when the chromosome in a genome changes in number.
However, mutations according to structural changes have four types, namely, deletions,
duplications, inversions and translocations. In deletion mutation occurs when a chromosome
losses it segment. In duplication, a duplicate of the chromosome segment is being added to the
original chromosome. Inversions are when an internal chromosome segment is detached and
inverted before reattachment. Inversion has two types, pericentric and the paracentric
inversions. Pericentric occurs when the centromere is included in the segment yet paracentric
occur when the centromere is not included in the inverted segment. Lastly, translocation
involves a exchange of segments between two non-homozygous chromosomes (Ramirez et al,
2013).
Gene mutation involves changes that are within one or more loci, which may involve one
or several nucleotides. Gene mutation is classified as Base pair substitution and Frameshift
mutation. Base pair mutations are because of copy errors in DNA replication (eg. a purine is
translated to another purine). A frameshift mutation is a genetic mutation caused by a deletion
or insertion in a DNA sequence that shifts the way the sequence is read (Ramirez et al, 2013).
The objectives for this experiment are as follows:
a. describe the different types of chromosomal mutations;
b. explain how chromosomal aberrations could affect the fertility of an organism;
c. explain the genetic consequences of chromosomal configurations at different
stages of meiosis;
d. describe the different classes of gene mutation;
e. explain the genetic consequences of the different classes of gene mutation
Methodology:
For the materials, we used 33 petri dishes,(2)500mL erlenmeyer flasks, (1)1000mL
beaker, (1)100mL graduated cylinder, (1) rubber band, (1) autoclavable bag, (1) pack cotton
buds, 1000mL distilled water and a microwave. For the agar preparation, we dissolved 21g of
Nutrient Agar (NA) in 700mL of distilled water. Then we placed the mixture inside a beaker.
After that we placed the beaker inside the microwave and set it to medium high for 3 mins. After,
we divided the mixture into two and place it onto two separate 500mL Erlenmeyer flasks. Then
we secured the flasks with the cotton plugs we made earlier.
*Computation:
33 plates x 20mL each plate = 660 or 700mL
X = 30.0g
700mL 1000mL
X = 21g of Nutrient Agar in 700mL dH20
In sterilization, we placed 33 petri dishes inside an autoclavable bag and secured it with a
rubber band. Then we wrapped the cotton buds with a few layers of foil. After, we covered the
flasks with foil, just enough to cover the cotton plug. Then we added distilled water onto the
pressure cooker. Then we waited for it to boil. Once the water inside boils, we placed the
materials inside the pressure cooker, and closed it tightly. Once we saw that there is already
steam coming out, we set the bulb to 15 psi. After that we waited until the pressure reached 15
psi. Once 15 psi is reached, we set the timer to 20 mins. After 20 mins, we turned off the
pressure cooker and waited until the pressure reaches zero. Then, we carefully removed the
cover of the pressure cooker. After all that we placed the petri dishes into the oven set at 50
degrees Celsius, and the flasks inside the refrigerator.
In the incubation and inoculation process of the experiment, we disinfected the laminar
flow to reduce bacterial contamination. Then, we placed the petri dish together with the agar in
Erlenmeyer flask inside the laminar flow. We heated the mouth of the Erlenmeyer flask
containing the nutrient agar and mouth of the petri dish prior to pouring the agar on the petri
dish. After pouring the agar in the 33 plates, we exposed it to UV light for 15 mins. Then we
heated the mouth of the beaker with the flame from the alcohol lamp. Then, we inoculate [Link]
in the culture media with a cotton swab. Then, we covered one of the two petri dishes with glass
and half of it with an index card. On the second petri dish, we covered half of it only with an
index card. After covering it we labelled it with exposed or not exposed and covered or not
covered. Then, we exposed it to UV light. Lastly, we took the plates and incubate them at 37
degrees Celsius for 24 hours.
After our experiment, we decontaminated the materials used. We placed the 33
petri dish, Erlenmeyer flask, and used cotton buds in an autoclavable bag and secure it with a
rubber band. Then we did the protocols that we did in sterilization.
Results and Discussion:
The pictures below are the Petri dishes with E. coli that are incubated at 37 degrees
Celsius for 24 hours. Figure 1 is covered with glass and index paper on the half of it while figure
2 is not covered with glass and the half is covered with index card.
Figure 1. Covered Figure 2. Uncovered
According to the results above, UV light isnt an effective agent of mutation because the
microorganism didnt grow on the exposed part of the petri dish. In figure 1, the bacteria had
growth because according to Dr. Cynthia Bailey glass can block UV rays but not that much. For
Figure 2, there is no growth in the exposed part of the petri dish because the UV is not filtered
out because there is no glass yet the index card blocked the UV rays. The paper provided a
good filter for the UV rays because its opaque color make the light bounce off. Overall, based
on my results the probability that an organism can crow under a UV light is 2 out of 10 colonies
because although it is covered with glass it is still possible for UV to pass through it. Using other
mutagens can make more accurate results. An example of a chemical mutagen is ethyl
methane sulfonate (EMS), methyl methane sulfonate (MMS), diethylsulfate (DES), and
nitrosoguanidine which de-acidify the acidic properties of DNA. They also have the property of
inducing error-prone DNA repair. This stimulation of error-prone repair allows all sorts of
mutation types to occur. Nitrous Oxide can also cause mutations in the DNA affecting protein
synthesis.
.
Conclusion:
I conclude that there are two classes of chromosomal mutation, by numerical and by
structure. The types of chromosome mutation by numerical changes are aneuploidy and
euploidy. Euploidy is when the whole sets of the chromosome change. Euploid is divided into
two sub class, namely, monoploidy and polyploidy. Monoploids contain only one set of
chromosomes while polyploids have more than two complete sets of chromosomes. Aneuploidy
is when the chromosome in a genome changes in number. It also divided into two types namely,
addition of chromosomes and loss of chromosomes. The addition of whole chromosomes group
is consist of the trisomics, tetrasomics and double trisomics. The loss of one or more
chromosome group is consist of the monosomics, nullisomics and double monosomics.
Moreover, the chromosomal mutations according to structural changes are deletion, duplication,
translocation and inversions. Deletion mutation occurs when a chromosome losses it segment.
Duplication happens when a duplicate of the chromosome segment is being added to the
original chromosome. Inversions are when an internal chromosome segment is detached and
inverted before reattachment. Inversion has two types, pericentric and the paracentric
inversions. Pericentric occurs when the centromere is included in the segment yet paracentric
occur when the centromere is not included in the inverted segment. Translocation involves a
exchange of segments between two non-homozygous chromosomes. The different classes of
gene mutation are base pair substitution and frameshift mutation. In base pair substitutions, the
nucleic acid is not translated correctly while in frameshift mutation a addition or deletion of a
nucleic acid or a codon results to change the way it should be translated. The genetic
consequence a base pair substitution and frameshift mutation is there will be changes in the
phenotype of an organism. UV rays can mutate an organism because photons of the UV are
absorbed by DNA molecules then an excited state is produced which allows for the
rearrangement of electrons resulting in the formation of photoproducts.