Genotypic and phenotypic analysis of

Arabidopsis thaliana S15aE r-protein genes

that contain insertional mutations

Stacey Abidayo
Research Headed by Dr. Szick-Miranda
I. Why Arabidopsis thaliana is an
Ideal Model for Analysis
 Genome sequenced
 Rapid life cycle
 Easily grown
 Many seeds
 Many mutants
 Genetic tools available
Arabidopsis thaliana
II. Importance of Ribosomes and Translation

 Translation is polypeptide synthesis

 Polypeptides are large molecules made of many
amino acids
 large polypeptides are called proteins
 Ribosomes consist of a large and small subunit
 Distinct structure allows translation to occur
III. Purpose of Our Research
 Group looking at a specific ribosomal protein gene,
S15a, which is considered to be part of the divergent
proteins
 Many genes encode this ribosomal protein gene and I’m
working with a specific one which is the S15aE
 Earlier research show that the S15a protein gene is part
of the cytosolic ribosome
 Research suggests that S15a is also incorporated into
the mitochondria
 Asking whether or not this gene is found in cytocol and
mitochondria or cytocol alone because there is evidence
that it is incorporated into the mitochondria
 Analyzing insertional mutations in genes
IV. Approach
 Looking at mutations that encode for those genes
that are available through the Salk Institute
Genome Analysis Laboratory
 We cannot assume that the seeds have an
insertional mutation without genotypic analysis
 Polymerase Chain Reaction (PCR)
 Electrophoresis
 Then we genotype to assure whether or not the
mutation that is Homozygous wild type,
Heterozygous, or homozygous mutation
 Hope to find a homozygous mutation then we can
phenotypically analyze how the mutation affects
the phenotype, its outward appearance of the
plant
V. Methods Utilized in the Lab
 DNA extractions
 PCR
 Electrophoresis
VI. DNA Extractions
 Purpose: Extract DNA
 Used mortar and pestle to grind
leaves of Arabidopsis thaliana to get
DNA from tissue
VII. Polymerase Chain Reaction
(PCR)
 Importance of primers
 Enables production of large quantities
of specified DNA
VIII. Electrophoresis
 Gel composed of agarose, buffer, and ethidium
bromide
 Agarose and buffer work as a matrix that allows DNA
to be separated
 Ethidium bromide binds to DNA which makes if
florescent when UV light is applied to it
 Purpose is to allow us to visualize the DNA
 Applying an electrical current to DNA, which is
negatively charged, pulls DNA towards the positive
end
 Distance traveled with relation to our size marker
helps estimate how many base pairs are in the strand
of DNA in which was extracted
Examples of Electrophoresis