Term
| These organisms lack a nucleus and any well defined organelles |
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Definition
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Term
| This kingdom include Archaebacteria and Eubacteria |
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Definition
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Term
| Organisms that have a membrane bound nucleus as well as defined organelles |
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Definition
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Term
| Not considered a living organism. obligatory intracellular parasites |
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Definition
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Term
| Each type of virus can infect and parasitize only a limited range of host cells |
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Definition
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Term
| bacteria reproduce through |
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Definition
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Term
| bacteria used for recombinant dna |
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Definition
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Term
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Definition
1. Non-pathogenic Strain 2. Cheap 3. Fast Growing 4. Genome Completely Sequenced |
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Term
| what are the 4 growth phases |
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Definition
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Term
This is a period of physiological adjustment involving the induction of new enzymes and the establishment of a proper intracellular environment. |
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Definition
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Term
This phase is characterized by maximal rates of cell division and increase in cell numbers and mass. This phase varies amount bacterial species – maybe as brief as about 20 minutes or as long as several hours (e.g. E.coli – 4 hours) |
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Definition
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Term
During this period, the essential nutrients begin to disappear, and there is a balance between cell growth and division and cell death. |
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Definition
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Term
During this phase, bacterial lysis and cell destruction cause a reduction in cell number. |
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Definition
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Term
| Used to carry and clone foreign DNA |
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Definition
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Term
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Definition
Extra chromosomal self replicating means of selection or marker restriction enzyme sites |
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Term
it is not part of the bacterial chromosome (Genome) |
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Definition
| vector characteristic: extra chromosomal |
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Term
it has its own Origin of Replication (Ori), so replication is independent of the Genome |
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Definition
| vector characteristic: self replicating |
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Term
a way of determining its presence in bacteria (e.g. antibiotic resistance) in the form of Marker Gene |
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Definition
| vector characteristic: means of selection or marker |
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Term
it has a site for inserting foreign DNA usually in the form of a Multiple Cloning Site (MCS) |
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Definition
| vector characteristic: restriction enzyme sites |
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Term
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Definition
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Term
small, double stranded (ds) circular DNA |
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Definition
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Term
viruses that infect bacteria (e.g. lambda phage) |
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Definition
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Term
combination of plasmid and phage sequences |
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Definition
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Term
| 2 plasmids we worked with |
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Definition
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Term
Site of where we insert the exogenous (foreign) DNA which have restriction enzyme recognition sequence sites |
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Definition
| Mulitple Cloning Site (MCS) |
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Term
| Gene for resistance to an antibiotic |
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Definition
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Term
| 2 methods used to transform E. coli (itroduce plasmid into the cell) |
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Definition
Electroporation method divalent cation method |
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Term
- electric current makes pores in the cellular membrane allowing the entry of the plasmid - very efficient, but very expensive |
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Definition
| electroporation transformation method |
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Term
- treating cells with compounds that contain divalent cations (e.g. calcium ions – Ca++) - not very efficient, but cheap without requiring expensive pieces of equipment |
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Definition
| Divalent cation transformation method |
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Term
1. Incubation on Ice “freezes” the membrane 2. Heat Shock fractures or distorts the membrane 3. Calcium ions coat the membrane (phospholipid bilayer) and the extraneous DNA masking the negative charges on the phosphate groups 4. Negatively charged DNA can now pass through the membrane into the cell |
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Definition
| Proposed mechanism of the divalent cation method |
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Term
| Why some colonies glow/flouresce under UV illumination |
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Definition
| they have pGlo; those that don't glow are pAMP |
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Term
A rapid, small scale method of obtaining or retrieving plasmid DNA (plasmid DNA + foreign/inserted DNA) from bacterial cells. |
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Definition
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Term
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Definition
Boiling method Alkaline Lysis method (we use) |
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Term
Goals using Alkaline Lysis Method
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Definition
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Term
| Goals using Sodium Acetate
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Definition
| To precipitate out proteins and bacterial DNA |
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Term
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Definition
| To precipitate out the plasmid DNA |
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Term
- ___ is a buffer that works at physiological pH (pH = 7.4). - ___ binds divalent cations in the lipid bilayer, thus weaking the cell membrane and also protecting DNA from DNAses. makes ___ |
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Definition
| Tris, EDTA, Glucose Tris EDTA (GTE) solution |
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Term
- This alkaline mixture lyses the bacterial cells. The detergent ___ dissolves the lipid component of the cell membrane (plasma membrane), as well as cellular proteins - The ___ denatures the chromosomal and plasmid DNA into single strands (ssDNA). The intact circle of plasmid DNA remain interwined. makes ___ |
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Definition
SDS, NaOH SDS/NaOH (Sodium dodyclsulfate + Sodium Hydroxide) - Alkaline Lysis Solution |
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Term
-The ___ returns the pH to neutral, allowing DNA strands to renature. - At the same time, the ____ precipitates the SDS from the cell suspension along with proteins and lipids with which it has associated. this makes ___ |
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Definition
Acetic acid, Sodium Acetate KOAc (Potassium Acetate + Acetic Acid): |
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