Secondary Treatment of H2O
~ involves the use of microbes to remove organic material from the liquid portion
of the sewage
Tertiary treatment
~ involves the removal of inorganic materials thru chemical means
Disinfection
~ disinfect the water to remove all microbes
Biological oxygen d...
✓~ refers to the use of living organisms to create products that are useful to humans
Recombinant DNA
✓~ DNA from two or more sources...allows for creation of DNA sequences not
otherwise found in biological organisms
Basic Mechanism of gene cloning
✓~ 1. isolate plasmid DNA from bacterial cell
2. isolate DNA of interest
3. Insert the gene of interest into plasmid DNA...forms recombinant DNA
4. insert the recombinant DNA into a bacterium
5. culture and grow the bacteria
,Restriction Enzymes
✓~ a type of endonuclease that cuts up DNA
*normally leave "sticky ends" of DNA
Restrictions sites (recognition sequences)
✓~ short nucleotide sequences that are recognized by restriction enzymes as the place
to cut DNA
normally palindromes
sticky ends
✓~ cut DNA that is left with short, single stranded sequences on both sides of the
fragments
DNA ligase
✓~ seals the two strands of "sticky ended" DNA together by catalyzing
phosphodiester bonds between the single-stranded breaks.
**results in recombinant DNA
,Electroporation
✓~ makes a cell competent to pick up DNA from the environment by applying an
electrical shock to it, which increases membrane permeability.
RecA proteins
✓~ competent cells are engineered to lack these proteins, which may try to otherwise
destroy the inserted sequence or attempt to correct the recombinant DNA
Reverse Transcriptase
✓~ turns single-stranded RNA into double-stranded DNA
it is an RNA-dependednt DNA polymerase
Complementary DNA (cDNA)
✓~ the DNA produced by reverse transcriptase and the RNA template
constructing cDNA
✓~ 1. apply s short poly-T primer to single-stranded RNA
2. add reverse transcriptase and 4 nucleotides
, 3. add RNaseH enzyme to cut up the RNA and regenerate the RNA primers
4. add DNA polymerase and DNA ligase to synthesize a new strand
cloning vector
✓~ provides a means for transferring a gene of interest to a host organism
"good" cloning vectors include
✓~ an origin of replication
a selectable marker
unique restriction sites (polylinkers or multicloning sites)
Plasmid
✓~ *most common cloning vector*
a self-replicating piece of extrachromosomal DNA found in prokaryotes
-mid-sized range (20,000 base pairs or less)
pBR322 and pUC19
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