Ch. 9 Biotech Flashcards
What is biotechnology
of microorganism to make a product
What is recombinant DNA?
r(DNA) insertion or modification of genes to produce wanted proteins through a vector for example
Vector
A DNA molecule that transports foreign DNA into a cell. The DNA molecule HAS to be self-replicating, such as a plasmid or viral genome.
Clone
clones of the cell that has the vector for the purposes of DNA
Examples of biotechnology?
They have done this for making bacteria when for insect eat plants they die, and for medications , or we can makee a genen that contains a degrative enzyme that canbe sued to clean up toxic waste, we can use it to make e coli make hGH humaan growth hormone
site-directed mutagenesis
targeted and specific change to a gene
Restriction enzymes
tools used for manipulation (we’ve exploited them to molecular scissors)
They cut specific sequences of DNA. They hydrolyze phage DNA in order to protect themselves, but they cannot kill themselves because they methylate some of their cytosines in their DNA
What restriction enzyme did we make from E.coli?
EcoR1
What is a blunt end?
When a restriction enzymes cuts at two places on the same strand
What is a sticky end
The end of the cut DNA by a restriction enzyme with some parts sticking out, these are better for rDNA because they introduce a plasmid that contains the complementary base paring
What is PCR?
Polymerase chain reaction (PCR) Used to make copies of DNA enzymatically. Clone DNA, amplify DNA number for detection. Diagnose genetic Disease and detect pathogens . Use heat to separate strands, then add the pieces needed to remake them primers, nucleotides, and DNA polymerase and make more, rinse and repeat
Why are shuttle Vectors important
They can survive in several different species, can be use to move cloned DNA to say bacteria to fungi etc.
Why can viral DNA be a useful vector
they can accept larger DNA than plasmids
What is the method used to shock DNA into a cell
Electroporation, used to insert DNA into a cell
What is the method called to introduce DNA into a cell via vectors
Transformation , they take DNA from the environment
Method where you enzymatically remove cell wall
Protoplast fusion, no cell wall means easier access. Then it can melt into another cell. this is useful for plants and algal cells
What is a gene gun for
Shoot DNA through the cellulose walls of plant cells, another method of DNA introduction
Method where you inject DNA into an animal cell
microinjection, because the needle is tiny
What is a genomic library
Collection f bacteria that each have different pieces of the genome of say an animals. Either in a phage (viral) library or plasmid library.
Why is eukaryotic DNA hard to clone genes from?
Has protein coding parts exons , and non protein coding parts introns, The introns make it big and bulky, and normally are taking out when converted to RNA, bacteria can’t always take them out themseslves. so an artificial gene is made that only contains exons through reveres transcriptase.
what is cDNA
Complimentary DNA that comes from mRNA, and has the introns and exons
What is synthetic DNA
made via DNA synthetic machines, much like typing an essay, but only up to 200 sequences and they need to be stitched together, and you need to know the instructions
What is a DNA library
collection of DNA seq that have been put into plasmids , So one colony would have a certain segment of DNA,
Why do we use E.coli a lot and what are disadvantages?
s easily grown and genomics are well known
But endotoxin must be removed from their outer cell wall. it causes fever and shock, and the cells need to be broken up in order to harvest proteins from there. it does not secrete them.
What other microbe is used in biotechnology and why?
Bacillus subtilis because it secretes protein. and are gram positive. Saccharomyces cerevisiae (yeast), its genome is 4 times bigger but it a well understood eukaryotic cell. they may be more successful in creating the product as well as continuously secreting them
What are mammalian cells used for
They can be used for making protein products for medical use because they secrete them and theres lower risk of toxins and allergies. For example CSF that stimulate the growth of cells that protect from infection. Normally we make little on our own.
What are plant cells used for
plants cells are altered by rDNA to make products like alkaloids (pain killers) rubber, sunscreen,. In order to modify them you also need bacteria sometimes.
what is a subunit vaccine
made through rDNA, a subunit vaccine only contains the protein part of a pathogen
Dna vaccines
made through rDNA, plasmids carry a viral gene under the control of a poromote active in human cells. its hard to get the plasmid DNA into human cells. Also we can use it for makeing human enzymes and hormones’
Gene therapy
rDNA allows for replacing defective genes or missing genes
Gene silencing
suppresses certain genes siRNA (small interefineg) bind to mRNA which is then destroyed by RISC thus silencing the expression of gene
Shotgun sequencing
A genome project where small pieces of genome in a living cell are sequenced and then assembled using a computer . Can be used to study microgorganisms that have not been cultured.
Human genome project
a project where they sequenced all og human DNA and hope it will provide info on diagnostics and treatments. Through blocking a gene we can determine its function (reverse genetics ). foward genetics is starting wwith pehnotype and looking for the genotype
What is it called determining the proteins made by a cell
proteomics
What is southern blotting
testing to see if a certain gene is present. can be used for forensics. band on the filter highlights the gene
Real-time PCR
newly made DNA is tagged with fluorescent dye. levels of florescence can be measured after every PCR cycle.
reverse transcription RT-PCR
is using reverse transcriptase by getting DNA from RNA or mRNA
How is it used in forensics
evidence collection, rape conviction ,tracing HIV , anthrax in mail etc. DNA fingerprinting can be used to identify bacterial or viral pathogenic
Why is agrobacterium important
it has a TI plasmid, that occurs naturally within it. as a result with rDNA we can make it pest resistant, herbicide resistant, suppressed gene that prevents enzymes from breaking down pectin, and thus have a longer shelf life or more nutritious
what are some ethical issues to look out for
accidental release, modified crops have to be safe for ppl and environment. Who has the righ tto have access to a persons genetic info?
