Definition
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Term
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Definition
| merlin holding two copies of chromosome segment |
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Term
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Definition
| complementation is a process in which a defective toy(gene) was traded in for a functional toy (gene) |
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Term
how do we move transposable elements, what are transposable elements?
what are thw two kinds of transposable elements in BACTERIA |
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Definition
transposable elements is a segment of DNA that moves from one spot to another DNA molecule
1. IS (insertion sequence )
2. transposom |
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Term
| insertion sequences (transposable elements) |
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Definition
| simplest transposable element- HAS NO ABX RESISTENCE |
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Term
TRANSPOSONS- abx resistance- has insertion sequences
TN5 and Tn10
transformers don't get sick (abx resistent) |
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Definition
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Term
| 2 mechanism for transposition |
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Definition
conservative
replicative
(conservative republicans, relocation) |
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Term
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Definition
transposon is cut out from one location and reinserted at a second location (TN5)
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Term
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Definition
| new copy of transposon produced and inserted at a second location (doubling effect |
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Term
| application of transposons |
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Definition
create mutants: key point. Tn5 insertion into a gene will cause the disruption of the gene. The purpose of Tn5 mutagenesis is to identify which gene is responsible for disease. For example, you isolated a pathogen from a diseased mouse, but you don't know which gene can produce toxin.
Anyway, after mutation with Tn5 transposon, there will be a lot of Tn5 mutants (maybe more than 5,000 mutants). So, let's assume there are 5,000 mutants and label 1 thru 5,000.
Now, let's apply Tn5 mutant #1 into a living host. If the host dies, mutation on gene #1 does not affect disease, because the disruption of the gene #1 is not important. The phenotype between the wild type (pathogen) and mutant #1 is the same.
However, when you **apply Tn5 mutant #2 into a living host, the host is alive. Disruption of gene #2 changes the phenotype, which means that gene #2 is responsible for causing disease. |
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Term
describe Molecular cloning |
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Definition
isolation and incorporation of a piece of DNA into a VECTOR so it can be replicated and manipulated |
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Term
| steps to molecular cloning |
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Definition
1. isolation and grag of dNA (isolate the donkey)
2. insert him into a cloning vector
a) vector- derived from plasmids or viruses
**b) DNA ligase- joinds dna molecule
3. Introduction of cloned DNA into host organism
a) transformation is usually used |
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Term
| example of an important cloning vector? |
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Definition
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Term
| important characteristic of plasmid cloning vectors |
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Definition
independent of orgin of replication
require selectable markers (abx markers) |
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Term
| describe what Blue/White screening is/ What function does it serve |
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Definition
blue- represents colonies which have no foreign DNA inserted into vector (no foreigners)
white- represents colonies with foreign DNA inserted |
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Term
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Definition
determinig the precise order of nucleotides in a DNA or RNA molecule
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Term
describe the sanger dideoxy method
function is to sequence DNA |
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Definition
oh --> no OH causes inhibition of DNA synthesis
REQUIRES OH
DIDEOXY= DNTPS prevents further DNA synthesis
so dNTP basically removes oh groups |
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Term
| primer used in sanger method has a nucleotide.... |
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Definition
| has nucletide complementary to 3 end of the region copied |
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Term
| addition of didribonuclease does what to DNA Replication process |
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Definition
didribonuclease stops replication
(did from dexter's labatory) |
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Term
| to sequence DNA what is required of the strand |
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Definition
| must be a single stranded DNA |
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Term
| after the transfer nucleotdoes in gel tubes how is the results read?? |
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Definition
| bottom to top, one base at a time. |
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Term
another method to sequence dNA is through pyrosequencing, and massively parallel liquid handling
describe these methods
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Definition
| - dna seq found by using light measuring tools, each dna attachment to template releases light, more atp more light |
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Term
LIGHT sequencing method: 3 steps
ppi + oxy-luci-fer-in =light
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Definition
1. you have a single stranded DNA
2. complementary nucleotodes are incorporated in strand
***1. 1 nucletode = 1 ppi
2. ATP sulfurylase converts PPi to ATP
***3. ATP drives luciferin --> oxyluciferin= LIGHT
ppi + oxy-luci-fer-in =light
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Term
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Definition
| CLOSED relies Solely on MANPOWER |
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Term
| function of polymerase chain reaction (PCR) in DNA replication |
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Definition
| method that produces multiple copies of DNA, causes exponential growth |
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Term
3 steps of polymerase chain reaction activity
describe the characteristcs of each step
DAE= fun pool days |
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Definition
1. denaturatio- 94 C
2. anealing- 50-60 C
3. extension- 92 C
"dogs are evil"
*know temperatures** |
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Term
Due to the high heat enviroment of DNA amplification, what is required for replication (2)
does it have proofreading activity |
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Definition
Taq polymerase- THERMOSTABLE DNA POLYMERASE
- has no proofreading activity
PFU- more thermostable than taq |
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Term
| does taq have proofreading activity |
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Definition
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Term
| which of the two themal polymerases posses proffreading activity |
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Definition
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Term
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Definition
1. leads to expoential growth
2. **valuable for cloning And sequencing
3. used to amplify DNA from mummified remains |
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Term
| which genes are the most abnundant class |
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Definition
METABOLIC GENES
mass=abundant |
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Term
| NUMBER OF TOTAL IDENTIFIED GENES |
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Definition
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Term
| DESCRIBE HYPOTHETICAL GENES |
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Definition
| UNCHARACTERIZED ORF'S PROTEINS THAT LIKELY EXIST BUT WHOSE FUNCTION IS UNKNOWN |
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Term
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Definition
| small solid state supports where fixed genes are spatially arranged in patterns |
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Term
function of microarray in experiments
5 functions |
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Definition
1.* global gene expression
2. expression of specific groups of genes under diff conditions
3.*expression of genes with unknown function; yield clues to possible roles
4.*comparison of gene content in closely related org
5.identification of specific organisms |
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Term
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Definition
| genome-widely study of the structure, function, and regulation of an organism's protein |
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Term
gene families, duplications, and deletions
paralogs |
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Definition
genes within an org is the result of gene duplication
(W/I species) |
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Term
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Definition
gnes found in one org similar to those in another org but differ in SPECIES
(other species***)
orth o log |
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Term
Evolution of virulence: pathogenicity islands
chromosomal island |
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Definition
| region of bacterial chrom of foreign orgin that contains clustered genes for some extra property such as virulence of symbiosis |
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Term
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Definition
| chromosomal islands containing genes for VIRULENCE; such as herbicidies |
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Term
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Definition
| chromosomal island contain genes for symbiosis duhh |
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Term
| what two components do bacteria cosist of? |
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Definition
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Term
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Definition
| shared by all strains of the sopecies |
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Term
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Definition
| includes extras in SOME not all species |
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Term
evolutionary analysis
major assumptions of molecular clocks (chronometers) |
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Definition
| nucletodies changes occur at constant rate, gen neural and random |
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Term
| most widely used molecular clocks |
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Definition
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Term
| what S rRNA is found in prok. |
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Definition
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Term
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Definition
18s rRNA
16 comes before 18, prok came before Euka |
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Term
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Definition
| basic local alignment search tool- genbank database helpful in identifiy gene sequences |
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Term
| classification of organism uses what kind of system? |
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Definition
| binomial system of nomenclature |
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Term
| directions for nomenclature |
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Definition
Genus species
G comes before S
1. genus first
2. species second
3. all Italic
4. First lefter Capitalized
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Term
Bacillus subtilis
Bacillus cereus
Escherichia coli K12
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Definition
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