Nate Rodriguez

10 Nov 2016
Genetics Lab
Yeast
Complementation testing is a technique used in genetics to determine the location
of alleles. It is based off the idea that multiple genes can affect a particular phenotype,
and these genes can be located on the same or different chromosomes. For example,
Organism A can have a genotype AAbb and will express the mutant phenotype.
Organism B can also express the same mutant phenotype but instead can have the
genotype aaBB. If these organisms were to mate and produce offspring, then every one
of their offspring could express the wild type phenotype or the offspring could express
the mutant phenotype. This technique is also sometimes referred to as cis-trans test. Cis
refers to the possibility that either both mutant alleles or both wild type alleles fall on the
same chromosome, and trans refers to the possibility that both mutant alleles and wild
type alleles fall on different chromosomes. The name cis-trans test gives better insight
into the idea of why two organisms that express the mutant phenotype can produce
offspring that express the wild type phenotype. If the mutant alleles are in a trans
configuration, then the offspring express the wild type phenotype. Furthermore, if the
alleles are in a trans configuration, then they are stated to complement each other.
However, sometimes the mutant alleles are in a cis configuration, which causes the
offspring to express the mutant phenotype just like the parents. The reason why a trans
configuration can produce a wild type phenotype is that with a trans configuration, the
offspring will contain a wild type allele and a mutant allele, and therefore, allows the

offspring to produce the necessary protein or enzyme required to express the wild type
phenotype. On the other hand, if the alleles are in a cis configuration, then the offspring
does not have the required genetic code to transcribe for the necessary enzyme or protein,
which allows the features of the wild type phenotype.
<https://www.britannica.com/topic/complementation-test>
While yeast is unicellular, it is still a eukaryote and is very important in the study
of cellular processes such as the reproductive and cell cycle. This is due to the fact that
its cellular activities are very similar to the cellular activities of species that are
considered to be more complicated. While yeast generally undergoes asexual
reproduction through the use of budding (a form of mitosis), it can also undergo sexual
reproduction just like other eukaryotic organisms. The benefit of sexual reproduction is
that yeast are able to transmit a lot of genetic diversity to their offspring. In order to
sexually reproduce, yeast must be a haploid cell, or only contain one set of genomic
material, and mate with another haploid cell. These haploid cells are called a and
(alpha). They are attracted to one another by the secretion of pheromones. Haploid cell a
secretes pheromone called a- factor and haploid cell secretes the pheromone - factor.
Once these pheromones are detected, it causes the cells to undergo a conformational
change and converge together. After the cells come into contact with each other, they can
begin a nuclear fusion, which allows them to form a zygote. With the formation of a
zygote, the yeast cells are initiated into a mitotic cycle, and the cells give rise to a diploid
bud and takes on the appearance of a dumbbell shape. Yeast then produces haploids
through the process of meiosis, which temporarily duplicates the cells genetic and divides
it evenly among 4 daughter cells. Then through the process of sporulation, haploid

spores are produced and are contained in a firm membrane structure called an ascus.
These spores are released once the environmental conditions are suitable and allow
optimal growth of yeast cells. Once released, the haploid a cells and the haploid cells
repeat the reproduction cycle.
Yeast can undergo a mutation that causes them to produce a red pigment and,
therefore, causes the colonies of yeast to have a red color instead of tan. This is caused
by a several different factors but the primary factor is the mutation in the DNA that
causes a disruption in the Adenine synthesis pathway. Normally, yeast cells produce
adenine through the creation of adenosine monophosphate or AMP. AMP is generally
made through the process of 12 enzymatic steps and starts with a sugar phosphate
compound called phosphoribosylpyrophate (PRPP) and is eventually attached to adenine,
the end product. However, just like any other cell, the process of making adenine can be
disrupted through the mutation of the ade1 and ade2 genes. When these genes become
disrupted, they allow a molecule called aminoimidazoleribotide (AIR) to accumulate in
the cytoplasm of the cell because an enzyme is no longer coded in the DNA of the yeast
cell. Then through the process of aerobic respiration, AIR is oxidized into a red pigment,
which then gives the colonies of yeast a red color. Any cellular reaction or genetic
mutation that inhibits either of these two processes, then the yeast maintains its white
phenotype. While, red yeast is still a viable phenotype and can survive and reproduce,
white yeast is considered to be dominant because it has a natural selection over red yeast
because it can grow better. Moreover, the red yeast mutant gives a lot of insight into the
study of the biosynthetic pathway of AMP production because it associates a certain
genotype, mutation in ade1 and/or ade2, with the formation of a certain phenotype, red.

After the mutation of red yeast, the next step would be to determine what
mutation took place and what gene did it affect in the yeast cellular DNA. Therefore, if
we obtain red yeast, then we received a mutation in the ade1 gene and/or the ade2 gene.
Furthermore, we more than likely also received other mutations throughout the DNA, but
none of them affect the aerobic growth in the red yeast cells.