Chapter 9 Flashcards
biotechnology
the use of microorganisms, cells, or cell components to make a product
example of products biotechnology
foods, antibiotics, vitamins, enzymes
recombinant DNA (rDNA) technology
the insertion or modification of genes to produce desired proteins
vector
self replicating DNA molecule used to transport foregin DNA into a cell
cells with vectors multiply to form
clone
clone
population of genetically identical cells arising from one cell; each carries the vector
selection
selecting for a naturally occurring microbe that produces a desired product
mutation
mutagens cause mutations that might result in a microbe with a desirable trait
site directed mutagenesis
a targeted and specific change in a gene
restriction enzyme cut specific sequences of
DNA
restriction enzymes _______ phosphodiester bonds between individual nucleotides at a specific recognition sequences on both strands DNA, and they are able to cut or digest the DNA sequence the same way every time
hydrolyze
restriction enzymes can occur
naturally
restriction enzymes typically cleave
unmethylated DNA
restriction enzymes create blunt ends or staggered cuts known as
sticky ends
___ ________ attaches the complementary sticky ends to one another this produces recombinant DNA molecule
DNA ligase
vectors carry new DNA to
desired cells
vectors must be able to
self replicate
______ and ______ can be used as vectors
plasmids, viruses
shuttle vectors
exist in several different species and can move cloned sequences among various organisms
PCR
polymerase chain reaction
PCR is a process of increasing small quants of DNA for
analysis
PCR is used for diagnostic tests for
genetic diseases and detecting pathogens
reverse transcription PCR uses ____ as a template and reverses the normal flow of genetic information. Utilizing the enzyme _______ ________ mRNA is used to create cDNA
mRNA, reverse transcriptase
what is cDNA
complementary DNA
the steps of PCR
- denaturation
- priming
- extension
denaturation PCR step is what temp
high temp
denaturation PCR results
DNA is incubated at high temp which causes the strands tp separate by breaking the hydrogen bonds
priming PCR temp
low temp
priming PCR results
DNA is incubated at a relatively low allowing primers to attach to the single stranded target DNA
extension PCR temp
intermediate temp
extension PCR results
DNA is incubated at an intermediate temp at which the DNA polymerase rapidly replicates DNA
PCR occurs in a machine called
thermocycler
PCR components
DNA polymerase, DNA primers, dATP, dCTP, dTTP, DGTP,
inserting foregin DNA into cells 3 types
transformation, electroporation, protoplast fusion
transformation
cells take up DNA from the surrounding environment
electroporation
electrical current forms pores in cell membranes
protoplast fusion
removing cell walls from 2 bacteria allows them to fuse
DNA can be inserted into a cell by 2 ways
Gene gun, microinjection
gene gun is used in
plants
microinjection is a technique that uses a _____ _______ with a diameter much smaller than the cell, and is able to puncture the plasma membrane, so that DNA can be injected through it
glass micropipette
genomic libraries are collections of clones containing different
DNA fragments
an organisms DNA is digested and spliced into _______ or _______ ______ and introduced into bacteria
plasmid, phage vectors
complementary DNA is made from mRNA by
reverse transcriptase
genomic libraries are used for obtaining eukaryotic genes because
eukaryotic DNA has introns that do not code for protein
mRNA has the _______ removed, coding only for the protein product
introns
synthetic DNA builds genes using a
DNA synthesis machine
in cloning it is necessary to select the particular cell that contains your specific ___ of interest
gene
blue white screening uses _______ _____ containing ________ resistance gene and b-galactosidase
plasmid vector, ampicillin
for blue white screening bacteria is grown in media containing ampicillin and X gal, a substrate for b- galactosidase, only the bacteria that picked up the plasmid will
grow
white colonies
bacteria that picked up the the recombinant plasmid will not hydrolyze lactose
blue colonies
bacteria with the intact lacZ gene will hydrolyze the x-gal and produce blue colonies
colony hybridization
common method of identifying cells that carry specific gene
colony hybridization uses DNA probes, what are these?
short segments of single stranded DNA complementary to the desired gene
e coli advantages for using it in making a gene product
easily grown and its genomics are unknown
e coli disadvantages for using it to make a gene product
produces endotoxins and does not secrete protein products
saccharomyces cereviase is what
bakers yeast
saccharomyces cereviase
- easily grown and has a ________ genome than bacteria
- expresses __________ genes easily
- likely to continuously secrete ________
larger, eukaryotic, product
plant cels and whole plants
- express ________ genes easily
- plants are easily grown, large scale and ___ cost
eukaryotic, low
mammalian cells
- express _________ genes easily
- can make products for ______ use
- ______ to grow
eukaryotic, medical, harder
subunit vaccines
made from pathogen proteins in genetically modified yeasts
DNA vaccines
usually _______ plasmid including a gene coding for a ____ protein
circular, viral
gene therapy
to replace defective or missing genes
gene editing uses
CRISPR
gene editing
to correct genetic mutations at specific locations
2 ways to conduct gene silencing
small interfering RNAs (siRNAs)
RNA interference (RNAi)
small interfering RNAs (siRNAs)
bind to mRNA which is then destroyed by RNA induced silencing complex
RNA interference (RNAi)
inserts DNA endowing siRNA into a plasmid and transferred into a cell
shotgun sequencing
sequences small pieces of genomes which are assembled by a computer
metagenomics
the study of genetic material directly from environmental samples
Human Genome Project
sequences the entire human genome
Human Proteome Project
will map proteins expressed in human cells
bioinformatics
understanding gene function via computer assisted analysis
proteomics
determining proteins expressed in a cell
reverse genetics
discovering gene function from a genetic sequence
southern blotting
DNA probes detect specific DNA fragments ________ which are restriction fragment length polymorphisms separated by ___ ________
RFLPs (rif lips), gel electrophoresis
genetic testing
screening of parental or fetal tissue for several hundred possible genetic diseases
DNA fingerprinting
is used to identify pathogens
PCR microarrays and DNA chips
can screen samples for multiple pathogens
forensic microbiology can be used to
find proper collection of evidence and establish a chain of custody
nanotechnology deals with the design and manufacture of extremely small electronic _______ and mechanical devices built at the ________ level of matter
devices, molecular
bacteria can make nanoparticles from
gold, silver, selenium and cadmium
Ti plasmid occurs in
agrobacterium tumefacines
Ti plasmid is a plasmid that can be used to introduce recombinant DNA into a
plant
Ti plasmid integrates into the plant genome and causes
tumor like growth
Bt toxins is normally produced by
Bacillus thuringiensis
Bt toxins can be produced at larger amounts by genetically modified
Pseudomonas fluorescens
Bt toxin is used as
pest control for insects that feed on crop plants
suppression of gene agricultural example
tomatoes that stay firm after harvest because the enzyme that breaks down pectin is suppressed
suppression of genes is achieves with
antisense DNA
antisense DNA is a length of DNA complementary to the ____ for the enzyme is synthesize and then is taken up by the cell where it binds to mRNA which inhibits _______ then the DNA-RNA hybrid is degraded by ______ in the cell and this frees up the antisense DNA to disable another mRNA
mRNA, translation, enzymes
when using DNA technology we need to avoid accidental release into the
environment
when making genetically modified crops we must make sure they are safe for
consumption and environment
