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83 Cards in this Set
- Front
- Back
what is the Central Dogma of Molecular Biology?
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DNA:Replication -> DNA:Transcription -> RNA:Translation ->
Protein |
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What is a gene?
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a nucleotide sequence in DNA to which a specific genetic function can be assigned.
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mRNA
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messenger RNA
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what does messenger RNA do?
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takes its genetic information or message out of the nucleus and into the cytoplasm of the cell.
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converts the nucleotide sequence of the mRNA into a specific sequence of amino acids to produce a specific protein.
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translation
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translation of mRNA is accomplished by translating a series of three nucleotides
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codon
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what molecules are involved in the central dogma of molecular biology?
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DNA RNA Proteins
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what are the processes involved in the central dogma of molecular biology?
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Replication, Transcription, translation
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what is a peptide bond?
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chemical bond between the carbonyl group of one amino acid and the amio group of a second amino acid (a special form of amide linkage)
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what are the start codons?
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met (AUG)
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what is the stop codons?
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UAA, UAG, UGA
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what amino acid is the start codon?
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Met
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what are restriction endonucleases?
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they are enzymes that catalyze the cleavage of the phosphdiester bonds within both strands of DNA.
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nucleotide sequence that is identical to its complementary strand when each is read in the same chemical direction
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palindromic sequences
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how are palindromes related to restriction enzymes?
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restricted enzymes only cut at very specific palindronic sequences of bases.
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what are "sticky" or "cohesive" ends?
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staggered cleavage site that can rejoin again.
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what are "blunt" ends?
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cuts that are opposite of each other.
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what is a recognitioon site where there are only four possible sequences?
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degenerate
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what are the recognition sites that are divided by a certain number of totally variable bases?
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hyphenated sites
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a powerful separation method frequently used to analyze DNA fragments generated by restriction enzymes
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agarose gel electrophoresis
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what does the gel consists of in a gel electrphoresis?
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microscopic pores that act as a molecular sieve.
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Since DNA has a strong _____ charge at neutral pH, it migrates through the gel towards the ____ electrode during electrophoresis.
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negative; postive
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the ____ the lnear fragment the faster it migrates in gel electrophoresis.
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smaller
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what was our source of DNA in gel electrophoresis?
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Lambda DNA
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Lambda phage DNA contains 10-16 base single-stranded regions at the 5'-3' terminus that are self-complimentary called___.
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cos ends
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what are our source of emzyme in gel electrophoresis? (6)
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Hind III, Hae III, Eco RI, Eco RII, Bgl I, Bam HI
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What is the restriction enzyme?
Bacillus globigii, Bacillus amyloliquefaciens H |
Bgl I; Bam HI
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What is the restriction enzyme?
Escherichia coli strain RY 13, Escherichia coli strain R 245 |
Eco RI; Eco RII
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What is the restriction enzyme?
Haemophilus segyptius, Haemophilus influenzae Rd |
Hae III, Hind III
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what is the horizontal axis in a restriction digest plot?
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migration distance
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what is the vertical axis in a restriction digest plot?
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log bases pairs
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how many amino acids are there?
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20
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the sequence or order of amino acids along a polupeptide chain is referred to as the ____ of the protein.
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primary structure.
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the spatial arrangement of the protein backbone that is generated from the folding of the polypeptide chian is called the _____ of the protein.
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secondary structure
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what is a central organizing feature of enzymes, antibodies, and most othe proteins?
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B-sheet
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the ____ of a potein describes the detailed features of the three dimensional conformation of the polypeptide chain.
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tertiay structure
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the ____ of a protein is due to the type and sequence of its constituent amino acids.
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three-dimensional structure
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the unfolding of a proteins conformation
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denauturation
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the movement of charges molecule under the influence of an electric feild.
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electrophoresis
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what are the difference between protein electrophoresis and DNA electrophoresis?
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DNA is (-) so it migrates to the (+) end of the gel. Amino Acids in Proteins are either (+) or (-) so they move in both directions.
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what is an iso-electric point?
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the pH at which an amino acid or protein does not migrated in an electric feild
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what is the role of pH of electrophoresis buffer n selarating proteins out of solution?
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it detemines the net charge on the protein molecules in the sample
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what will happen if you increase or decrease the pH of the buffer?
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the higher the charge the faster the protein will travel. so if the protein's pH is close to the pH buffer it will move slower. if the pH of the protein is farther from the pH buffer it will move faster.
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what type of buffer was used in protein electrophoresis?
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electrophoresis buffer
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what is genetic transformation?
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change caused by genes and it involves the insertion of a gene(s) into an organism in order to change the orgamism's traits
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who normally do genetic transformation?
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agriculture, bio-remediation, and medicine
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what is the procedure of genetic transformation?
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reqires two steps:
1. a calcium cholorid buffer which offsets the environment to neutralize the similar charges 2. heat shock which increase the permeability of the bacterial membrane |
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what is your vessel for ntroducing genes?
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plasmid (pGLO)
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what must be done to your organism to get this vessel in?
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heat shock to break bacterial membrane
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provides a solid matrix to support bacterial growth
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agar
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a carbodrate, normally used as source of food by bacteria
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arabinose
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a protein the provides resistance to the antibiotic amplicillin
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beta-lactamase
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appluing bilogy in the real world by the specific maipulation of living organisma to produce potentially beneficial products
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biotechnology
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when a population of cells is prepared by growth from a single cell, all the cells in the poulation will be geneticlly identical
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cloning
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a clump of genetically identical acterial cells groowing on an agar plate.
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colony
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the liquid and solid media are referred to as LB broth and agar are made from an extract of yeast and an enzymatic digest of meat byproducts the provides a misture wich are nutrients for bacterial growth.
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culture media
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the manilulation of an oganism's genetic material by introducing or eleminating specific genes.
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genetic engineering
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polymerizes when heated to form a solid gel and functons to provide a solid support on which to culture the bacteria
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agar
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the unique 3-D conformation cuases it to resonate when exposed to ultraviolet and give off energy in the form of visible green light.
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green fluorescent protein
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a circular DNA molecule, capable of autonomus replication, carrying one or more genes for antibiotic resistance proteins and a cloned foreign gene such as GFP
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plasmid
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plasmid containing the GFP sequence and ampicillin resistance gene which codes for beta-lactamase
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pGLO
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the process of cutting and recombining DNA fragments as a means to isolate genes or to alter their structure and function
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recombinant DNA technology
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process of iddentifying wanted bacteria from a bacterial library
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screening
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minimizing the possibility of outside bacterial contamination during an experiment through observance of cleanliness and using careful labroatory techniques
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sterile technique
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process of passing an inoculation loop with bacteria on it across an agar plate
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streaking
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an autonomously replicating DNA molecule into which foreign DNA fragments are inserted and then paropagated in a host cell
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vector
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what plasma did we use for transformation lab
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pGLO
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what genes did pGLO contain?
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GFP and a gene for resistance to the antibiotic ampicillin.
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why did we use E. coli in transformation lab?
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Because it is a bacteria that reproduces fast.
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what specific E. coli did we use in transformation lab and why?
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E. coli HB 101 because it is non-pathogenic, non-infectious, and only grows in the presence of cetain media
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what is a phenotype?
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the observable character of a cell or organism
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what is amp?
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ampicillin: we put ampicillin into the plates to kill off the bacteria that doesnt have the GFP gene or the antibiotic gene. (if doesnt have antibiotic gene then doesnt have GFP gene so we dont need them)
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what is LB?
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luria-bertani: which is the liquid or solid media where the nutrients neeed to make bacteria grow is located
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what is ara?
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arabinose: which turns on the GFP in transformed cells
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the liquid that is present above the pellet is called____
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supernatant
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HIC
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hydrophobic interaction chromatography
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when the lasate is applied to the column, the hydrophoi proteins that are applied the column in a high salt buffer will stick to the beads while all other proteins in the mixture will pass through. when the salt is decrease, the hyfrophobic proteins will no longer stick to the beased and will drop out the botton of the column in a purified form
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HIC
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spinning a mixture at very high speed to separate heavy and light particles
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centrifuge
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what is the result of centrifuge?
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a pellet and supernatant
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what did the pellet in the expression lab contain?
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bacteria
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what happens to GFP in each buffer? (Binding, Wash, and Elution)
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Binding: the GTP changes its comfomation to where the hydrophobic parts are on the outside sticking to the beads.
Wash: the weaker hydrophobic parts drop through the column Elution: the GFP returns to its original conformation and drops through the column. |
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enzyme needed to break open bacteria cell walls
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lysozyme
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a process for separating complex liquid mistures of proteins or other molecules by passing a liquid misture over a column containing a solid mixture.
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chromatography
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