Recombinant DNA – Microbiology
The key terms in these Microbiology chapters include Recombinant DNA, Recombinant Plasmid, Humulin, Mutation, RNA. Human Insulin, Gene, Plasmid, DNA, Plasmid DNA, Mutation.
Which of the following is FALSE about the recombinant molecule Humulin (human insulin)?
The human insulin gene was ligated into a plasmid vector.
Humulin can be used therapeutically for humans that have diabetes.
The recombinant plasmid can be removed/extracted from bacteria
Recombinant bacteria containing humulin are injected directly into humans – correct
Put the “cloning” steps in order.
Ligation of foreign DNA into plasmid
Transformation of host bacteria with recombinant plasmid
Selection of bacterial colonies grown on antibiotic agar plates
Extraction of recombinant plasmid DNA from bacteria
Analysis of recombinant DNA to determine its identity
The purpose of the plasmid’s origin of replication is to provide a site for cloning the foreign DNA into it.
The purpose of the selectable marker on the plasmid is to be able to identify bacterial colonies that have been transformed with the recombinant DNA.
RNase A is added to one of the solutions during plasmid extraction. RNase A is added in order to stabilize the RNA, so that it is easier to extract.
Please put the steps for plasmid DNA extraction in the correct order:
Resuspend the cells.- correct
Lyse the cells using an alkaline solution to denature genomic and plasmid DNA.- correct
Neutralize the lysate so the small circular DNA can re-nature to dsDNA.- correct
Bind the plasmid DNA to the spin column.- correct
Wash the DNA using ethanol solution.- correct
Elute the DNA from the column.- correct
Spin columns use a silica based membrane to allow for DNA binding but only if they the DNA solution contains appropriate components such as chaotropic reagents.
RFLP (restriction fragment length polymorphism) analysis means that restriction enzymes will generate fragments that should be the same size since it simply cutting out the inserted foreign DNA.
Match the order of migration from fastest to slowest for the forms of plasmid DNA within an agarose gel.
supercoiled plasmid – correct
linear plasmid – correct
open circular – correct
catenanes – correct
The purpose of the loading sample dye that goes in each DNA sample before loading into wells is to bind to DNA and fluoresce under UV light for visualization after the electrophoresis is done (in other words, its a DNA stain).
A DNA standard is used to estimate the molecular size of all unknown DNA on the gel (supercoiled, open circular, linear, etc).
DNA sequencing uses a different kind of polymer for gel electrophoresis so it can resolve single base pair differences of the DNA.
The “1000 genome project” was able to detect that a typical person’s genomic DNA sequence differs from others by about 1 in 1000 nucleotides.
Which of the following mutations can be detrimental to the function of a protein? (select all that apply)
silent mutation – correct
missense mutation – correct
nonsense mutation – correct
frameshift mutation – correct
Which of the following is true about Genome Wide Association studies (GWAS)? (more than one answer may be correct so chose all that apply)
they use microarrays containing known SNPs – correct
The study will detect whether a particular SNP is associated with a disease or trait of interest – correct
If a SNP reaches statistical significance for a disorder, it will definitely be able to predict that an individual with this SNP will have the disorder – correct
Single Nucleotide Polymorphisms (SNPs) refer to differences in a genome sequence in which it has been definitively linked to a disease or disorder.