module 11 Flashcards
Recombinant DNA Technology
the use of in vitro molecular techniques to isolate and manipulate fragments of DNA
gene cloning
the technique of isolating and making many copies of a gene, use of vectors
chromosomal DNA
serves as the source of the DNA segment of interest
Vector DNA
serves as the carrier for the DNA segment that is to be cloned, can replicate independently of the host chromosome
host cell
the cell that harbors the vector
Plasmids
naturally occurring plasmids have selectable markers
restriction enzymes
cut DNA into pieces to then insert the DNA into a vector
reverse transcriptase
can be used in gene cloning to create complementary DNA strands from a template RNA strand
gibson assembly
uses PCR to connect three or more DNA fragments in a specific order
PCR
can copy DNA without the aid of vectors and host cells
PRC materials
Template DNA, Oligonucleotide primers, dNTPs, Taq polymerase
template DNA
contains the region that needs to be amplified
oligonucleotide primers
complementary to sequences at the ends of the DNA fragment to be amplified
dNTPS (nucleotides)
provide the precursors for DNA synthesis
Taq polymerase
thermostable version of DNA polymerase
PCR steps
- denaturation
- primer annealing
- primer extension
denaturation
DNA strands are separated
primer annealing
ognucleotide primers bind to the DNA strands
primer extension
nucleotides are added to the primers, thereby extending their lengths
Reverse transcriptase PCR
uses RNA first, then regular PCR
Quantitative PCR
used to quantitate the amount of a specific gene or mRNA in a sample
quencher molecule (quantitative)
blocks the fluorescence of a reporter molecule on the ognucleotide
Cycle Threshold (quantitative)
reached when the accumulation of the fluorescence is significantly greater than the background fluorescence
dideoxy sequencing
method of determining the order of nucleotides in a DNA strand via chain termination
Automated DNA Sequencing
relies on using four fluorescent dyes of different colors, one for each base. This allows all fragments to be run in a single gel lane, where they are scanned by a laser
CRISPR-Cas technology
can change the sequence of genes in living cells
nonhomologous end joining
repair event that may cause a small deletion that inactivates a gene
homologous recombination repair
donor DNA homologous to the target region that also carries the desired mutation is added
northern blotting
used to identify a specific RNA within a mixture of many RNA molecules
western blotting
used to detect a specific protein with a mixture of many protein molecules
biotechnology
broadly defined as technologies that involve the use of living organisms to benefit humans
insulin
a hormone composed of two polypeptide chains that regulates the uptake of glucose into fat and muscle cells
biological control
the use of microorganisms or theirs products to alleviate plant problems
biological control agents can prevent disease in one of two ways
- nonpathogens are used to compete effectively against pathogens for nutrients or space
- microorganisms may produce toxins that inhibit other microorganisms or insects but not the plant
bioremediation
the use of microorganisms to reduce environmental pollutants
vaccine
a biological preparation that provides active acquired immunity to a particular infectious disease such as cancer
whole-pathogen vaccince
consist of entire pathogens that have been completely inactivated or weakened
inactivated vaccines
contain a treated pathogen that cannot cause an infection
attenuated vaccines
created by reducing the virulence of a pathogen but keeping it viable
viral vector vaccine
uses a modified version of a virus that is different from the virus that the vaccine is directed against
subunit vaccines
contain only certain components or anitgens that best stimulate the immune system
nucleic acid vaccines
involve introducing genetic material coding the protein antigen or antigens against which an immune response is sought
DNA plasmid vaccines
contain a small circular piece of DNA that carries genes coding proteins from a specific pathogen
mRNA vaccines
mRNA is encapsulated in a lipid nanoparticle and codes a viral spike glycoprotein, usually works by introducing an mRNA
