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BIO 219 Questions With Accurate Solutions (Pass!)
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BIO 219 Questions With Accurate Solutions (Pass!)
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BIO 219 Questions With Accurate Solutions (Pass!)Shotgun Cloning Right Ans - Generate random fragments of genomic DNA
-different sizes
-can optimize conditions to generate larger fragments
Fragments will contain Right Ans - regulatory elements and the coding
sequences
Once inserted in a vector Right Ans - Each vector contains a separate slice
of the genome
whole genome is represented
Stable library of the entire genome to go back to again and again
Shotgun Cloning Right Ans - Pick a genome (A. ficsheri)
Pick a vector
Pick a restriction enzyme
How long is the lux operon Right Ans - 8.5 kb long
A. fischeri Right Ans - Bacteria found throughout marine environments
Auto-induced bioluminescence Right Ans - An operon that allows A.
fischeri to glow under the right conditions
Vector types Right Ans - Plasmids
Phagemids
Bacteriophage Vectors
Cosmid Vectors
Bacteria/Yeast artificial chromosomes
Phagemids Right Ans - phage + plasmid
-package ssDNA in presence of a helper phage
Bacteriophage Vectors Right Ans - Based on the lambda virus
larger capacity >25 kb per virus
, Cosmid Vectors Right Ans - lambda cos sites + plasmid
->35 kb (35-45kb) inserts availabe
-can be packed in viral proteins for infectivity
-low copy number in bacteria
Bacterial/yeast artificial chromosomes Right Ans - -large, low copy number
1-2 per cell
->300 kb inserts are acceptable
-Good for sequencing
We chose phagemids Right Ans - because it will operate as a plasmid
its small circular DNA
-1-200kb
-can accept DNa fragments from 1-10kb
-extra-chromosomal
Self replicating
Exist in nature to carry antibiotic resistance and other survival genes
can be transmitted between two bacteria
been engineered into excellent vectors
What makes a good plasmid vector? Right Ans - Size
-large enough to hold workable quantities of foreign DNA
-Small enough to be retained by the host
-small enough to be able to distinguish from host chDNA
High copy number
-50 to 100 per cell
Origin of replication (ORI)
- Must be recognizable by the host's replication machinery
- Must be able to copy often and independently of host DNA
Multiple Cloning site
-region of dna containing recognition sequences for many restriction enzymes
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