Term
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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 |
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Term
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Definition
| circular pieces of DNA that replicate independently of the host's chromosomal DNA |
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Term
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Definition
mainly bacteria, but can also exist in...
-archaea
-eukaryotes |
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Term
| some eukaryotes that can have plasmids |
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Definition
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Term
| some beneficial functions plasmids can provide to the host |
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Definition
-resistance to antibiotics
-degradative functions
-virulence |
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Term
| what natural plasmids contain |
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Definition
-origin of replication (all have this)
-typically include a gene that is advantageous for survival |
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Term
| what artificial plasmids used in the lab are used for |
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Definition
| to introduce foreign DNA into the cell |
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Term
| minimally, plasmids made in the lab contain... |
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Definition
-origin of replication
-selection marker
-cloning site |
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Term
| what makes plasmids attractive tools for life scientists and bioengineers? |
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Definition
| The ease of modifying plasmids and the ability of plasmids to self-replicate within a cell |
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Term
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Definition
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Term
| some vector elements within a plasmid |
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Definition
-Origin of Replication (ORI)
-Antibiotic Resistance Gene
-Multiple Cloning Site (MCS)
-Insert
-Promoter Region
-Selectable Marker
-Primer Binding Site |
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Term
| Origin of Replication (ORI) |
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Definition
| DNA sequence which allows initiation of replication within a plasmid by recruiting transcriptional machinery proteins |
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Term
| Antibiotic Resistance Gene |
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Definition
| Allows for selection of plasmid-containing bacteria. |
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Term
| Multiple Cloning Site (MCS) |
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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. |
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Term
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Definition
| Gene, promoter or other DNA fragment cloned into the MCS for further study. |
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Term
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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. |
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Term
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Definition
| The antibiotic resistance gene allows for selection in bacteria. However, many plasmids also have selectable markers for use in other cell types. |
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Term
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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. |
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Term
| Due to their aritificial nature, lab plasmids are commonly referred to as... |
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Definition
| “vectors” or “constructs” |
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Term
| what scientists may do to insert a gene of interest into a vector |
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Definition
| utilize one of a variety of cloning methods (Restriction enzyme, Ligation indepenent, Gateway, Gibson, etc) |
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Term
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Definition
-Restriction enzyme
-Ligation indepenent
-Gateway
-Gibson
-etc. |
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Term
| how the cloning method is chosen |
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Definition
| based on the plasmid you want to clone into |
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Term
| Addgene’s Plasmid Guide includes information about... |
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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 |
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Term
| Generally, scientists use plasmids to... |
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Definition
| manipulate gene expression in target cells |
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Term
| some characteristics of plasmids that enable molecular biologists to broadly utilize plasmids across a wide range of applications |
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Definition
-flexibility
-versatility
-safety
-cost-effectiveness |
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Term
| Some common plasmid types |
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Definition
-Cloning plasmids
-Expression plasmids
-Gene knock-down plasmids
-Reporter plasmids
-Viral plasmids
-Genome engineering plasmids |
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Term
| some things plasmids are being used for |
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Definition
vectors for experiments encompassing...
-fluorescent imaging
-recombinant DNA technology
-mass protein production
-disease modeling
-drug discovery
-genome editing |
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