Genetics 4

DNA Sequencing Primer Site
Allows for identification of the DNA segment cloned into the plasmid

Multiple cloning site
Regions wjere pieces of DNA are cloned into the plasmid

Origin of replication
Enables replication of the plasmid within the bacterial cell

lacZ gene
Allows for selection of. Arterial cells that contain a cloned DNA fragment

Ampicillin resistance gene (amp R)
Allows for the selection of bacterial cells that contain the cloning plasmid

The products of restriction gives room can be visualized by gel electrophoresis, which separates fragments based on their size.

True or false?

True

A 1.5kb fragment of DNA is cloned into a plasmid vector that is 5.5kb long at the EcoRI site, and the plasmid vector is then used to transform bacteria.

If the plasmid DNA is then extracted from a single bacterial colony and digested with EcoRI, what digestion products will be produced if the plasmid contains the fragment?

One 1.5kb fragment and one 5.5kb fragment.

The role of primers in PCR is
To define the target region and provide a 3′ end that can be extended by taq polymerase.

If there are five molecules of DNA containing the target region at the beginning of a PCR reaction, how many copies of the target will be present after three rounds of amplification?
40

5*2=10
10*2=20
20*2=40

Immediately after the primers have annealed to the target sequence,
The temperature is raised so that taq polymerase can extend the primers

Cloning
The assembly of assembling and copying recombinant DNA fragments

Restrictive enzymes
Cut DNA in specific places to get DNA fragments

Vector
A DNA molecule able to replicate when introduced into host cells

DNA library
Collection of cloned DNA

PCR
Is a method used to quickly amplify DNA in vitro without using hosts and vectors

Southern Blotting
Desperation of DNA fragments by gel electrophoresis and hybridization with labeled probes. It shows the presence of specific sequences and fragment sizes

DNA sequencing
Used to determine the primary structure of DNA

Gene Knockout
Can be created to terminate the expression of a gene

Recombination can be used for
Achieving gene knockout by swapping the gene of interest with a copy that will produce a non functional protein

Transgenic
Organisms that can be created to express inserted gene of interest, which may come from a different species

How are ddNTPs structurally different from dNTPs?
ddNTPs lack a 3′ hydroxyl group

How does this structural difference in ddNTPs make it valuable in a chain termination of DNA sequencing?
Once they are incorporated with DNA, they terminate the chain elongation, thereby allowing multiple fragments of DNA to be formed and analyzed to reveal the base sequence of original DNA

Which type of DNA library represents the genes expressed. Y a given cell at a certain time!
cDNA

What is not used to make a cDNA library?
RNA Polymerase

Which type of library should be screened to isolate the promoter of a gene?
Genomic library

What is a probe in molecular biology?
A DNA or RNA molecule used in hybridization reactions

Northern boots are used to study what type of molecule?
RNA

Which is most often the first step in characterizing a cloned DNA
Restriction mapping

Why are filters overlaid with x ray film when a cDNA library is screened?
To visualize probe hybridization events

Some proteins require posttranslational modification for proper folding and function. Which host cells would enable the production of a human protein that requires this modification?
Yeast

Prokaryotic genomes are always much smaller than eukaryotic genomes.
False

What is not a common feature of eukaryotic genomes?
Operons

The study of orthologs would be useful to determine the function of a specific gene in a species.
True

Globin superfamily diversity arose primarily from
Gene duplication and divergence

What info can be obtained from hybridization micro arrays?
The expression level of genes

Which technique would be used if a proteomics approach were used to find the cause of a muscle disorder?
Determining which environmental factors influence the expression of the protein of interest

Two dimensional gel electrophoresis is used in proteomics to
Separate different proteins

What percentage of human genome codes for proteins?
2%

Which program in the human genome project was designed to ensure that personal genetic info would not be used in discriminatory ways?
ELSI