Many regions of non-coding eukaryotic DNA previously thought to be "junk" are now known to contain sequence elements important to regulating gene expression. What approach can be used to identify important non-coding regulatory regions when annotating a newly sequenced genome? O comparing CDNA to genomic DNA to validate that the gene is expressed O identifying restriction enzyme recognition sequences in the genome O phylogenetic footprinting to identify conserved non-coding sequences O searching for start/stop codons and splice recognition sites that predict where a gene might be located

Many regions of non-coding eukaryotic DNA previously thought to be "junk" are now known to contain sequence elements important to regulating gene expression. What approach can be used to identify important non-coding regulatory regions when annotating a newly sequenced genome? O comparing CDNA to genomic DNA to validate that the gene is expressed O identifying restriction enzyme recognition sequences in the genome O phylogenetic footprinting to identify conserved non-coding sequences O searching for start/stop codons and splice recognition sites that predict where a gene might be located

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter13: An Introduction To Genetic Technology

Section: Chapter Questions

Problem 8QP

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In generating mutations in a bacterial gene involved in antibiotic resistance, a number of point mutations are isolated that render the bacteria sensitive to the antibiotic. You would like to sequence the gene in order to characterize the mutations, but unfortunately, your lab partner just finished the last of the lab's supply of DNA polymerase. The only things at your disposal are materials for performing a western blot, allowing you to visualize the protein encoded by the gene. How would you identify which mutations are likely to be the result of a missense mutation, which are likely to be the result of a nonsense mutation, and which are likely to be the result of a frameshift mutation?
a To make a genomic library useful for sequencingan entire genome, why would you ordinarily fragment the genomic DNA by mechanical shearingforces like sonication rather than by cutting theDNA with a restriction enzyme?b. Suppose that you wanted to make a genomic library to determine the complete sequence of anewly discovered organism’s genome, but you didnot have a sonicator readily available. Explain howyou could nonetheless use two or more restrictionenzymes to make libraries whose clones could besequenced so that a computer could assemble thegenomic sequence.c. Suppose you only had a single restriction enzymeavailable, and you want to make a single genomiclibrary from which you could assemble the genomic sequence. How might you be able to achievethis goal? (Hint: See Problem 9.) To make this library, would it be preferable to use a restriction enzyme that recognizes a 4-base, 6-base, or 8-basesequence of DNA?
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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.
a. What sequence information about a gene is lackingin a cDNA library?b. Can clones in a cDNA library contain 5′ UTR sequences? 3′ UTR sequences?c. Would you be likely to find on average longerORFs in cloned sequences from a genomic libraryor from a cDNA library? Explain
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Consider a partial restriction digestion, in which genomic DNA is exposed to a small, limiting amount ofa restriction enzyme for a very short period of time.a. Would the resultant fragments be longer or shorteror the same size as those produced by a completedigestion?b. If you prepared genomic DNA from a tissue sample containing millions of cells, would the fragments produced by partial digestion of DNA fromall of these cells be the same or different?