UF MCB 3020 Exam 3 UPDATED
Questions and CORRECT Answers
biotechnology - CORRECT ANSWER - refers to the use of living organisms to create
products that are useful to humans
Recombinant DNA - CORRECT ANSWER - DNA from two or more sources...allows for
creation of DNA sequences not otherwise found in biological organisms
Basic Mechanism of gene cloning - CORRECT ANSWER - 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 - CORRECT ANSWER - a type of endonuclease that cuts up DNA
*normally leave "sticky ends" of DNA
Restrictions sites (recognition sequences) - CORRECT ANSWER - short nucleotide
sequences that are recognized by restriction enzymes as the place to cut DNA
normally palindromes
sticky ends - CORRECT ANSWER - cut DNA that is left with short, single stranded
sequences on both sides of the fragments
DNA ligase - CORRECT ANSWER - seals the two strands of "sticky ended" DNA
together by catalyzing phosphodiester bonds between the single-stranded breaks.
**results in recombinant DNA
,Electroporation - CORRECT ANSWER - makes a cell competent to pick up DNA from
the environment by applying an electrical shock to it, which increases membrane permeability.
RecA proteins - CORRECT ANSWER - 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 - CORRECT ANSWER - turns single-stranded RNA into double-
stranded DNA
it is an RNA-dependednt DNA polymerase
Complementary DNA (cDNA) - CORRECT ANSWER - the DNA produced by reverse
transcriptase and the RNA template
constructing cDNA - CORRECT ANSWER - 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 - CORRECT ANSWER - provides a means for transferring a gene of
interest to a host organism
"good" cloning vectors include - CORRECT ANSWER - an origin of replication
a selectable marker
unique restriction sites (polylinkers or multicloning sites)
Plasmid - CORRECT ANSWER - *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
bacteriophage - CORRECT ANSWER - virus that infects bacteria
carry smaller fragments between 9,000-25,00 base pairs
cosmid - CORRECT ANSWER - a hybrid between a plasmid and a phage
carry large fragments between 30,00 and 47,000
artificial chromosomes - CORRECT ANSWER - synthetic chromosomes that contain
fragments of DNA integrated into a host chromosome
1. P1 artificial chromosomes (PACs)
2. Bacterial artificial chromosomes (BACs)
3. yeast artificial chromosomes (YACs)
**carry up to 1 MILLION bp
DNA analysis - CORRECT ANSWER - used to determine the location of genes,
differences in genes, and where and when genes are expressed
Includes...
1. PCR
2. gel electrophorese
3. restriction fragment analysis
4. blotting techniques
Polymerase chain reaction (PCR) - CORRECT ANSWER - quickly and easily makes
copies of even a very small amount of DNA.
targets the DNA with the following:
1. Primers
2. thermostable DNA polymerase (taq polymerase)
3.deoxyribonucleotide triphostates (dATP, dCTP, dGTP, dTTP)
Questions and CORRECT Answers
biotechnology - CORRECT ANSWER - refers to the use of living organisms to create
products that are useful to humans
Recombinant DNA - CORRECT ANSWER - DNA from two or more sources...allows for
creation of DNA sequences not otherwise found in biological organisms
Basic Mechanism of gene cloning - CORRECT ANSWER - 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 - CORRECT ANSWER - a type of endonuclease that cuts up DNA
*normally leave "sticky ends" of DNA
Restrictions sites (recognition sequences) - CORRECT ANSWER - short nucleotide
sequences that are recognized by restriction enzymes as the place to cut DNA
normally palindromes
sticky ends - CORRECT ANSWER - cut DNA that is left with short, single stranded
sequences on both sides of the fragments
DNA ligase - CORRECT ANSWER - seals the two strands of "sticky ended" DNA
together by catalyzing phosphodiester bonds between the single-stranded breaks.
**results in recombinant DNA
,Electroporation - CORRECT ANSWER - makes a cell competent to pick up DNA from
the environment by applying an electrical shock to it, which increases membrane permeability.
RecA proteins - CORRECT ANSWER - 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 - CORRECT ANSWER - turns single-stranded RNA into double-
stranded DNA
it is an RNA-dependednt DNA polymerase
Complementary DNA (cDNA) - CORRECT ANSWER - the DNA produced by reverse
transcriptase and the RNA template
constructing cDNA - CORRECT ANSWER - 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 - CORRECT ANSWER - provides a means for transferring a gene of
interest to a host organism
"good" cloning vectors include - CORRECT ANSWER - an origin of replication
a selectable marker
unique restriction sites (polylinkers or multicloning sites)
Plasmid - CORRECT ANSWER - *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
bacteriophage - CORRECT ANSWER - virus that infects bacteria
carry smaller fragments between 9,000-25,00 base pairs
cosmid - CORRECT ANSWER - a hybrid between a plasmid and a phage
carry large fragments between 30,00 and 47,000
artificial chromosomes - CORRECT ANSWER - synthetic chromosomes that contain
fragments of DNA integrated into a host chromosome
1. P1 artificial chromosomes (PACs)
2. Bacterial artificial chromosomes (BACs)
3. yeast artificial chromosomes (YACs)
**carry up to 1 MILLION bp
DNA analysis - CORRECT ANSWER - used to determine the location of genes,
differences in genes, and where and when genes are expressed
Includes...
1. PCR
2. gel electrophorese
3. restriction fragment analysis
4. blotting techniques
Polymerase chain reaction (PCR) - CORRECT ANSWER - quickly and easily makes
copies of even a very small amount of DNA.
targets the DNA with the following:
1. Primers
2. thermostable DNA polymerase (taq polymerase)
3.deoxyribonucleotide triphostates (dATP, dCTP, dGTP, dTTP)
