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BIOL 3110L Quiz 2
BIOL 3110L Quiz 2
27
Biology
Undergraduate 3
01/20/2018

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Term
how PCR is done
Definition
1: near-boiling temperature causes double-stranded DNA to separate, or denature into single strands
2: temperature is decreased and short DNA sequences known as primers bind, or anneal, to complementary matches on the target DNA sequence. The primers bracket the target sequence to be copied
3: At a slightly higher temperature, Taq polymerase binds to the primed sequences and adds nucleotides to extend the second strand. This completes the first cycle.
4: repeat 1-3
Term
plasmid
Definition
circular pieces of DNA that replicate independently of the host's chromosomal DNA
Term
where plasmids are found
Definition
mainly bacteria, but can also exist in...
-archaea
-eukaryotes
Term
some eukaryotes that can have plasmids
Definition
-yeasts
-plants
Term
some beneficial functions plasmids can provide to the host
Definition
-resistance to antibiotics
-degradative functions
-virulence
Term
what natural plasmids contain
Definition
-origin of replication (all have this)
-typically include a gene that is advantageous for survival
Term
what artificial plasmids used in the lab are used for
Definition
to introduce foreign DNA into the cell
Term
minimally, plasmids made in the lab contain...
Definition
-origin of replication
-selection marker
-cloning site
Term
what makes plasmids attractive tools for life scientists and bioengineers?
Definition
The ease of modifying plasmids and the ability of plasmids to self-replicate within a cell
Term
anatomy of a plasmid
Definition
[image]
Term
some vector elements within a plasmid
Definition
-Origin of Replication (ORI)
-Antibiotic Resistance Gene
-Multiple Cloning Site (MCS)
-Insert
-Promoter Region
-Selectable Marker
-Primer Binding Site
Term
Origin of Replication (ORI)
Definition
DNA sequence which allows initiation of replication within a plasmid by recruiting transcriptional machinery proteins
Term
Antibiotic Resistance Gene
Definition
Allows for selection of plasmid-containing bacteria.
Term
Multiple Cloning Site (MCS)
Definition
Short segment of DNA which contains several restriction sites allowing for the easy insertion of DNA. In expression plasmids, the MCS is often downstream from a promoter.
Term
Insert
Definition
Gene, promoter or other DNA fragment cloned into the MCS for further study.
Term
Promoter Region
Definition
Drives transcription of the target gene. Vital component for expression vectors: determines which cell types the gene is expressed in and amount of recombinant protein obtained.
Term
Selectable Marker
Definition
The antibiotic resistance gene allows for selection in bacteria. However, many plasmids also have selectable markers for use in other cell types.
Term
Primer Binding Site
Definition
A short single-stranded DNA sequence used as an initiation point for PCR amplification or sequencing. Primers can be exploited for sequence verification of plasmids.
Term
Due to their aritificial nature, lab plasmids are commonly referred to as...
Definition
“vectors” or “constructs”
Term
what scientists may do to insert a gene of interest into a vector
Definition
utilize one of a variety of cloning methods (Restriction enzyme, Ligation indepenent, Gateway, Gibson, etc)
Term
some cloning methods
Definition
-Restriction enzyme
-Ligation indepenent
-Gateway
-Gibson
-etc.
Term
how the cloning method is chosen
Definition
based on the plasmid you want to clone into
Term
Addgene’s Plasmid Guide includes information about...
Definition
-molecular cloning
-how to choose a plasmid vector
-molecular biology tools and references
-how to maintain your plasmid stocks
-multiple protocols and troubleshooting tips to make plasmid usage as simple and straightforward as possible
Term
Generally, scientists use plasmids to...
Definition
manipulate gene expression in target cells
Term
some characteristics of plasmids that enable molecular biologists to broadly utilize plasmids across a wide range of applications
Definition
-flexibility
-versatility
-safety
-cost-effectiveness
Term
Some common plasmid types
Definition
-Cloning plasmids
-Expression plasmids
-Gene knock-down plasmids
-Reporter plasmids
-Viral plasmids
-Genome engineering plasmids
Term
some things plasmids are being used for
Definition
vectors for experiments encompassing...
-fluorescent imaging
-recombinant DNA technology
-mass protein production
-disease modeling
-drug discovery
-genome editing
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