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Processing and Translation
May 26 Class 11
43
Biology
Undergraduate 2
05/26/2015

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Cards

Term

What is the ribosome made up of?

X

Definition

two subunits

3 sites for binding rna a,t

 

Term
What binds small subunits of mRNA in the initiation process of translation in the ribosome?
Definition
Ribosome
Term
Describe the ribosome binding in eukaryotes (Translation: Initiation)
Definition

Binds to 5 cap and slides alond mRNA until it gets to first good start codon

can be one start site on eukaryotic mRNA

Eukaryotic mitochondria (an organelle) and viruses have different systems

Term
Describe the ribosome binding in eukaryotes (Translation: Initiation)
Definition
Term
What is a polycistronic message?
Definition
multiple separate coding swquences on the same RNA
Term
Where does the start codon line up at?
Definition
P-site
Term
What binds to start the start codon?
Definition
Initiator tRNA
Term
Where does the large subunit join?
Definition
the complex
Term
What is the binds the large subunit to the P site?
Definition
Initiator tRNA
Term
What energy does the large subunit use to join the complex?
Definition
GTP
Term
What are the three steps of Elongation?
Definition

Codon recognition

Peptide bond formation

Translocation

Term
Discibe Codon recognition
Definition

occurs in the Aminoacyl site- tRNA loaded into A site

tRNA anticodon base pairs with mRNA codon- correct tRNA selected by sampling

Protein factor that loads tRNA requires energy-GTP

Term
What kind of energy does Codon Recognition for Protein factors that load tRNA?
Definition
GTP
Term
What occurs during peptide bond formation?
Definition

Peptide bond is formed between peptide (short smino acid chain) on tRNA in P-site and aminoacyl tRNA in A-site

Breifly tansfers growing peptide to the A-side

(connects amino acid coming in and releases the chain)

Term
What occurs during Translocation?
Definition

mRNA/tRNA complex is moved within the ribosome to the next site

requires energy GTP

tRNA in A-site, with peptide, ends up in P-site

Empty tRNA in P-site ends up in E-site=free to exit

Term
What happens after all three steps in elongation occur?
Definition
A new codon has been lined up in A-site, ready for a new tRNA to repeat the cycle
Term
Do all three steps in Elongation require energy?
Definition
No peptide bond formation doesn't
Term
What occurs in termination?
Definition

Protein release factore (RFs) bind to stop codon in A-site

Hydrolyzes bond connecting peptide to P-site tRNA- releases completed protein from ribosomes

Ribosomes dissociates, releasing mRNA- requires energy GTP

Term
When does GTP hydrolysis occur in intiation?
Definition
Term
When does GTP hydrolysis occur in elongation?
Definition
Term
Describe polyribosomes (polysomes)
Definition

Multiple ribosomes can translate the same mRNA at once

-you don't have to wait for one to finish before then next starts- as soon as one clears the start codon, another can load on and immediately start

Allows rapid translation

Term
What is only seen in bacteria?
Definition
linked transcription and translation
Term
In the bacteria, describe linked transcription and translation
Definition

They are not segregated in time and space

Multiple RNA polymerases can be transcribing the same gene at once

Polyribosomes can form on each mRNA

This results in a whole lot of protein being made from the same gene, all at once

Term
The basic ideas of transcription and translation are universal. Give and example of this
Definition
genes from one species can be transferred to another and still work
Term
What is a mutation?
Definition
Change in DNA sequence
Term
How are mutations classified?
Definition
The effect they have on the protein from the gene that has been mutated
Term
What is a silent mutation?
Definition

where there is no effect in an aminoacid that is coded by new codon as old

one nucleotide-pair is changed but the amino acid is still the same

Term
What is a missense mutation?
Definition

changes amino acid

Different amino acid coded by new codon

changing one nucleotide pair which changes the whole amino acid

Term
What is a nonsense mutation?
Definition

Prematurely terminates protein

New codon is stor codon

protein product is frequently completely non-functional and the mRNA is targeted for destruction

Term
What is a frameshift mutation?
Definition

insert or delete bases

3 diff ways to divide a swquence into codons, start establishes which will be used, take out or add bases you shift into a diff reading freame, changes all subsequent amino acids

Term
What does the frameshift usually lead to?
Definition

Usuaully terminatess early

Tandom chance will get a stop codon 3/64 or 1/20

analogous to a nonsense mutation

Term
What does the Addition or removal of a number of bases divisible by 3 - entire codon (s) do?
Definition
adds or removes amino acid(s) but doesn't change remainder of protein
Term
How do mutations arise?
Definition
errors in DNA replicatipn
Isomerization of bases during DNA replication
Induced changes-UV light, Ionizing Radiation, Chemical mutagens
Term
What is protein targeting?
Definition
◦ Signal sequence - targets protein to endomembrane system
 Specific sequence found of first amino acids of peptide
emerging from ribosome
 Recognized by signal recognition protein (SRP)
 Entire ribosome dragged to rough ER
 Injects growing protein into or through ER membrane
 Signal peptide removed from mature protein
Term
What is recombinant DNA?
Definition
contains DNA from multiple sources- recombined
Term
What is a plasmid and what does is do?
Definition
Circular extra-chromosomal DNA molecules
 Like the genome, they have a single origin of replication
 Can be gained and lost by their host
 Can carry genes for specific functions not encoded by
the genome
 Can be made to carry
foreign DNA
 Used for genetic
engineering cloning
Term
What are 3 restriction on enzymes?
Definition
isolated on bacteria
cleave DNA at specific sequences
Any restriction fragments with compatible ends can be glued together by ligase to produce new combinations
Term
How is being isolated from bacteria a restriction to enzymes?
Definition
They use them to protect
from and scavenge foreign
DNA (e.g. viruses)
Term
How is cleaving DNA at specific sequences a restriction to enzymes?
Definition
 “Restriction sites”
 Leave single-stranded
“sticky ends” with unique
self-complementary
sequences
Term
What is gel electrophoresis?
Definition
Separate nucleic acids (DNA, RNA)
or protein from each other, based
on size
Term
Describe how gel electrophoresis works
Definition
 Electrical current drives molecules
forward through a gel, (a Jello-like
block of carbohydrate or synthetic
molecular chains that form a grid,
allowing selective passage of
molecules based on size
 Results in size-based separation
 Shorter molecules travel fastest/farthest
 Visualized using stain
 Bands can then be cut out as isolated
pieces
Term
What is Recombinant DNA?
Definition
Isolate individual pieces of DNA
 Cut with restriction enzymes
 Separate by size on gel
◦ Ligate isolated pieces together to form recombinant plasmid
◦ Transform into bacteria
◦ Grow a lot of bacteria, letting bacteria make a lot of copies
of the plasmid
Term
Why make recombinant DNA?
Definition
Make a lot of pieces of specific DNA
 For subsequent genetic manipulation or study
 Have bacteria make a lot of a specific protein
 To change the characteristics of the host
 To study the protein
 For drugs
Analogous approaches
can be used to genetically
engineer eukaryotes
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