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Questions and Answers
What will occur when a dsDNA fragment with EcoRI sites is treated with the EcoRI enzyme?
What will occur when a dsDNA fragment with EcoRI sites is treated with the EcoRI enzyme?
In the forward orientation of the DNA fragment, which of the following is true regarding the coding and template strands?
In the forward orientation of the DNA fragment, which of the following is true regarding the coding and template strands?
What would be the result of aligning the sticky ends of the DNA fragment in reverse orientation?
What would be the result of aligning the sticky ends of the DNA fragment in reverse orientation?
Which of the following scripts the correct amino acid sequence for the protein initiated from the given mRNA in the forward orientation?
Which of the following scripts the correct amino acid sequence for the protein initiated from the given mRNA in the forward orientation?
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What might be the consequence of having an incorrect orientation when ligating the dsDNA fragment into a cloning vector?
What might be the consequence of having an incorrect orientation when ligating the dsDNA fragment into a cloning vector?
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Study Notes
Double Stranded DNA Orientation
- Double-stranded DNA (dsDNA) fragments have a 'forward' and 'reverse' orientation.
- The coding region of the DNA fragment, is flanked by restriction sites.
- Restriction enzyme EcoRI creates a 5' overhang.
- A cloning vector cut with the same enzyme can create a matching, complementary overhang to allow for a ligation.
- The orientation of the DNA fragment dictates whether a protein can be synthesized.
Forward Orientation
- In forward orientation, the sticky ends of the DNA fragment align correctly to synthesize a protein.
- mRNA is produced based on this orientation.
- Example mRNA sequence and corresponding protein are given.
Reverse Orientation
- In reverse orientation, the sticky ends of the DNA fragment do not align correctly for protein synthesis.
- mRNA is not properly formed for protein production .
- Example is provided of a mRNA sequence that produces no protein in the reverse direction
- Assuming the promoter is to the left of the restriction site, the top strand is the coding strand, and the bottom strand is the template strand.
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Description
This quiz covers the orientations of double-stranded DNA (dsDNA) fragments, specifically focusing on forward and reverse orientations. Understand how these orientations affect mRNA production and protein synthesis, including the role of restriction enzymes like EcoRI. Examples will illustrate the differences in functionality between correct and incorrect alignments.
