The sequence at one end of one strand of the Drosophilatransposon Mariner is shown below (dots indicatesequences within the transposon):5′ TTAGTTTGGCAAATATCTCCCTTCCGCCTTTTTGATCTTATGT... 3′You obtain a mutant bacterial strain tagged with anengineered Mariner transposon, cut the genomicDNA from this strain with the restriction enzymeMboI (whose recognition site is ^GATC), and circularize the resultant DNA fragments by diluting therestriction enzyme digest and adding DNA ligase.a. Design two 17 bp PCR primers that you could useto identify (by inverse PCR) the gene into whichthe transposon inserted.b. What DNA sequence will be amplified from thecircularized fragments of the mutant genome?Show the extent of this DNA sequence on a mapof the genome of the mutant strain, indicating thelocations of the transposon insertion and any relevant sites for the enzyme MboI.
Bacterial Genomics
The study of the morphological, physiological, and evolutionary aspects of the bacterial genome is referred to as bacterial genomics. This subdisciplinary field aids in understanding how genes are assembled into genomes. Further, bacterial or microbial genomics has helped researchers in understanding the pathogenicity of bacteria and other microbes.
Transformation Experiment in Bacteria
In the discovery of genetic material, the experiment conducted by Frederick Griffith on Streptococcus pneumonia proved to be a stepping stone.
Plasmids and Vectors
The DNA molecule that exists in a circular shape and is smaller in size which is capable of its replication is called Plasmids. In other words, it is called extra-chromosomal plasmid DNA. Vectors are the molecule which is capable of carrying genetic material which can be transferred into another cell and further carry out replication and expression. Plasmids can act as vectors.
The sequence at one end of one strand of the Drosophila
transposon Mariner is shown below (dots indicate
sequences within the transposon):
5′ TTAGTTTGGCAAATATCTCCCTTCCGCCTTTTTGATCTTATGT... 3′
You obtain a mutant bacterial strain tagged with an
engineered Mariner transposon, cut the genomic
DNA from this strain with the restriction enzyme
MboI (whose recognition site is ^GATC), and circularize the resultant DNA fragments by diluting the
restriction enzyme digest and adding DNA ligase.
a. Design two 17 bp PCR primers that you could use
to identify (by inverse PCR) the gene into which
the transposon inserted.
b. What DNA sequence will be amplified from the
circularized fragments of the mutant genome?
Show the extent of this DNA sequence on a map
of the genome of the mutant strain, indicating the
locations of the transposon insertion and any relevant sites for the enzyme MboI.
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