Isolation of Plasmid DNA Exam Questions and Answers Latest Update 2025
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Isolation of Plasmid DNA
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Isolation Of Plasmid DNA
Isolation of Plasmid DNA Exam Questions and Answers Latest Update 2025
what are plasmids? - Answers Plasmids are extrachromosomal elements that exist outside the chromosome, but within the bacterial cell.
They replicate in sync with what? - Answers They replicate in sync with the chromosome (term...
Isolation of Plasmid DNA Exam Questions and Answers Latest Update 2025
what are plasmids? - Answers Plasmids are extrachromosomal elements that exist outside the
chromosome, but within the bacterial cell.
They replicate in sync with what? - Answers They replicate in sync with the chromosome (termed
stringent replication) and so are partitioned to, and therefore inherited by, all daughter cells.
They may be transferred to the same or closely related bacterial species, by what process? - Answers by
the process of conjugation (by way of a mating tube that connects two cells).
plasmids can be transformed into bacterial cells, artificially or naturally? - Answers They may also be
transformed into bacterial cells, artificially, as naked DNA, as we do in molecular biology.
how do plasmids replicate? - Answers Plasmids replicate from a fixed origin of replication, with
bidirectional 'rolling circle' forks moving clockwise and anticlockwise around the double-stranded
plasmid until they meet on the other side, whereupon the replicating enzyme complexes disengage,
enabling the new daughter plasmids to separate.
what do the "new" plasmids comprise? - Answers the "new" plasmids comprise one original parental
strand (the template) and one newly copied complementary daughter strand.
Large native plasmids have been engineered into much smaller what? - Answers "cloning vectors"
how many copies of plasmids per cell in molecular cloning? - Answers Copies per cell may be 10 to 50,
but may be amplified to >1000 copies with chloramphenicol (relaxed replication control)
what is the size range of plasmid? - Answers Size range of 2 to 6 kb
what was the first engineered (artificial) plasmid to be employed as a cloning vector in molecular
biology? - Answers pBR322
The origin of replication allows the plasmid to what? - Answers The origin of replication allows the
plasmid to be copied independently of the bacterial host chromosome.
what is a recombinant plasmid? - Answers cloning vectors are used to insert foreign genes, resulting in
the formation of a recombinant (hybrid, or chimeric) plasmid.
what is the crucial point of cloning? - Answers The crucial point of cloning is the insertion of foreign DNA
into a cloning vector.
why is cloning is the insertion of foreign DNA into a cloning vector a crucial point? - Answers In this way,
the resulting recombinant plasmid "perpetuates" the gene independently of its original host and enables
the gene to be processed, manipulated, and studied by a number of methods and technologies,
including: gene mapping, DNA sequencing, sub-cloning, amplification of copy number, site-directed
mutagenesis.
, the smaller the plasmid, the, what? - Answers meaning that it will generate a higher copy number per
bacterial cell - and therefore a higher yield of plasmid DNA
smaller the plasmid, the higher copy number per bacterial cell generated, why? - Answers It has only
one antibiotic resistance gene for selection and maintenance in a bacterial culture.
what is LacZ? - Answers a gene promoter
where is LacZ located? - Answers inside the MCS, and the lacI repressor gene, together constitute the
gene expression system for this and most other commercially available expression vectors.
what constitues the gene expression system for this and most other commercially available expression
vectors? - Answers LacZ and the lacI repressor gene.
the expression system allows what? - Answers This expression system allows a cloned gene to be held
'silent' (not expressed at all), or to be switched on (expressed) at different rates (like the force of water
from a tap).
This expression system allows a cloned gene to be held 'silent' (not expressed at all), or to be switched
on (expressed) at different rates (like the force of water from a tap), why? - Answers The overall
intention is to produce a protein product from the cloned gene
what is an important feature of this plasmid - Answers The last important feature of this plasmid (and
other modern vectors) is the MCS (Multi-Cloning Site) region that contains an engineered short
sequence of DNA with many restriction enzyme sites in tandem as indicated.
The partner plasmid to pUC19, namely pUC18, is identical in all respects except for what? - Answers The
partner plasmid to pUC19, namely pUC18, is identical in all respects except for the orientation of the
MCS.
- Answers Isolating plasmids requires the selective removal of all other macromolecular components
(polysaccharides; chromosomal DNA; RNA; proteins; lipids; membrane fragments
Isolating plasmids requires what? - Answers Isolating plasmids requires the selective removal of all other
macromolecular components (polysaccharides; chromosomal DNA; RNA; proteins; lipids; membrane
fragments)
what are the major macromolecular types? - Answers polysaccharides; chromosomal DNA; RNA;
proteins; lipids; membrane fragments, of which there are four in the cell cytoplasm and two in the cell
membrane.
In some bacteria, what is also present in significant quantity? - Answers polysaccharides
There are three major considerations in nucleic acid isolation and purification procedures, what are
they? - Answers (1) How to maximize the yield of plasmid DNA (by amplification); (2) How to isolate and
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