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Biotechnology Recombinant DNA
quiz 7 unfinished
the use of microorganisms, cells ro cell compoennts to make a product
ex. foods, antibiotics, enzymes, proteins, vitamins
Genetic engineering and Recombinant DNA technolgoy
insertion of genes in an organism to produce desired protesin/enzymes or creat new cells that produce chemicals that an organism doesn't naturally make.
the culturing of a naturally-occurring microbe that produces a desired product.
1. Artificial selection
survival of the fittest in biotechn.
selectiion by humans of organisms with desirable characteristics.
mutagens cause mutations that might result in a microbe with a desirable trait.
2. Random selection
Mutation - Site-directed mutagenesis
changing a specific gene in DNA to change a protein
Mutation - random selection
select and culture microbe with the desired mutation.
specific sequences of DNA
Naturally occur in some species of bacteria. ex. to protect against incoming DNA to cut it up before it cuases any damage.
- tells restriction enzymes not to cut its own DNA
different Restriction enzymes
cuts are staggered, producing "
- become single strands
if two fragments of DNA from different sources have been cut with the same restriction enzyme, the two fragments will have
complimentroy sticky ends
and can recombine.
will then covalently link the sugar-phosphate backbones, producing a
recombinant DNA molecule
Transport DNA into the desired cell
ex. Plasmids, transposons, and viruses
Importanc characterists of Vectors
Should resist destruction by the recipient cell
vectors are highly resilient
b. Viral DNA more likely to remain intact.
Should carry a
, which makes it easy to retrieve clones containing the vector.
are plasmids capable of existing in several different species, and are used to move genes from one species to another.
Gene therapy and vectors
Vectors used to insert functional genes into human cells that have defective genes.
Polymerase Chain Reaction (PCR)
used to make multiple copies of a piece of DNA.
PCR is used to:
DNA for recombination
DNA to detectable levles
- to determain order of letters
Diangose genetic disease
Steps of PCR:
1.Original piece of DNA placed in a test tube with DNA primers, Individual DNA necluotides, DNA polymerease. Than test tube placed in a
for 1 min to
seperate the strands
of the original piece of DNA
3. Temp lowered
C for 1 min. -
DNA primers attach to the individual DNA strands
4. Temp increased to
for 1 min -
DNA polymerease synthesized a complimentory strand of DNA
5. Cycle is repeated many times, resulting in an exponential
increase in the number of DNA pieces
, all are identical to original DNA
6. DNA polymerase taken from the thermophilic bacterium "
" becuase of high temps.
Ways a recombinant DNA can be inserted into a cell
cell treated with chemicals to make them "competent" or able to take up external DNA. Ex. CaCl2, than heat shocked
applied an electrical current that forms small pores in the plasma membrane so the DNA can pass through.
To take up DNA better by those cell that have cell walls they need to be converted to protoplasts. (G + doesn't have cell walls, so thye take up DNA beter)
Protoplasts put into a solution with added
that increases the rate of fusion of two cells so that the 2 chromosomes can undergo natural recombination and become one cell.
pariticles of gold are coated with DNA and shot out of a gne gun witha burst of helium so that it penetrates plant cell walls - doen to eukaryotes - they can survive better.
a glass micropipette is used to inject DNA into a cell - humans
are made up of
of an entire genome stored in bacterial plasmids or in phages.
made by a DNA synthesis machine - only short pieces
Cloning the genes of Eukaryotes
) is made from an mRNA template by the enzyme
The result is a DNA molecules without introns.
This is used to obtain eukaryotic genes that can then be inserted into a prokaryotes.
Selecting a clone
from a gene library that contains your gene of interests
1. Blue-white screening
2. Colony hybridization
selects for clones that have any recombinant DNA
: identifies the clone with your gene of interest.
DNA probes are short sequences of single-stranded DNA that are compoimentary to the desired gene.
a. DNA probes are synthesized in the labe
b. Some DNA probes contain fluorescent dye
c. Some DNA probes contain radiocative phosphorous
DNA probes will bind with the gene of interest and serve as indicators