DNA Technology Flashcards
cut dsDNA at specific sites
Restriction endonucleases
vehicles that carry DNA fragments into cells
vector
phage mediated gene transfer into a prokaryotic cell
Transduction
non-phage mediated gene transfer into a prokaryotic cell
Transformation
Repairs single strand breaks in vivo and dsDNA breaks in vitro
DNA ligase
Gene transfer into a eukaryotic cell
Transfection
dsDNA added to blunt ends to make them sticky ends
Linker
A vector with an incorporated gene
Recombinant DNA molecule
Method used to identify DNA fragments
Southern blotting
Method used to identify RNA fragments
Northern blotting
Identification of proteins using specific antibodies
Western blotting
DNA is cut with a restriction enzyme and run on gel electrophoresis. This method is used during Southern Blotting
RFLP - restriction fragment length polymorphism
Restriction enzymes are used during what blotting method?
Southern
A genomic library containing the entire genome of an organism cloned into bacterial colonies.
DNA library
libraries include complementary DNA sequences produced using RT of all the mRNA expressed in a tissue. These DO NOT contain introns.
cDNA library (complementary)
Joining of a dideoxynucleotide will stop DNA polymerase at that base, with the type of base known at that position. Lining up all the tubes adjacent on gel electrophoresis will produce a number of varying fragment lengths. What is this method?
Sanger (dideoxy) method.
Remember: read from smallest fragment (BOTTOM) to largest fragment (TOP)
different flourolabel are attached to each ddNTP. All four reactions are performed in one tube and electrophoresis performed using 1 lane. The sequence is read as the light produced by the labeled bands is read by a detector. What is this method?
Automated DNA sequencing
This method is Useful in diagnosing genetic disease, identifying carriers, detecting infectious organisms, DNA profiling.
PCR
point mutation in sickle-cell beta-globin removes a MstII restriction site in the gene, producing a larger fragment. This is an example of what test method?
Southern blotting
amplification of the mutated region produces bands that are the desired length in repeat.
PCR analysis
What type of testing would you perform on a patient with Myotonic Dystrophy whose DNA segments are >4kb?
Southern blotting
*PCR cannot be used for this because the DNA segments are too large to migrate.
Patients with hemophilia A are deficient in what available recombinant protein?
Factor VIII
Patients with hemophilia B are deficient in what available recombinant protein?
Factor IX
Therapy where one cures or prevents disease by modifying an individual’s genes or their expression
Gene therapy
Therapy where alterations of genes in somatic cells are used to treat a specific disorder
Somatic therapy
Therapy where alteration of genes in gametes are used to prevent disease in child
Germline therapy
Replacing a mutated gene with a healthy one
Gene replacement therapy
Introduction of a new gene in order to treat disease
Gene therapy for non-inherited diseases
Inactivation of a mutated gene that is functioning improperly
Gene-blocking therapy
What is the most common vector used for gene therapy?
Adenovirus
Therapy where pt’s cells are removed from the body, treated, and re-introduced.
Ex vivo gene therapy
SCID, Gaucher Disease, and Malignant melanoma are treated using what kind of gene therapy?
Ex vivo gene therapy
Circulating cells are treated with this kind of gene therapy?
In vivo gene therapy
Cystic fibrosis, various cancers, Hemophilia B are all treated using what type of gene therapy?
In vivo gene therapy
Hepatocytes and skin fibroblasts are treated with this gene therapy to integrate a gene to promote the production of Factor IX. What is the therapy type and disease associated?
In vivo somatic gene therapy for Hemophilia B.
