Imagine you want to study human crystallins, proteins present in the lens of the eye. To obtain a sufficient amount of the protein, you decide to clone the crystallin gene. 1. Would you construct a genomic library or a cDNA library? (2 pts) 2. Explain the difference between these two types of molecular libraries. (2 pts) 3. What material would you use as a source of DNA or RNA? (2 pts)
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Would you construct a genomic library or a cDNA library? (2 pts)** To clone a gene for protein expression (to obtain a sufficient amount of the protein), a cDNA library is generally preferred. This is because cDNA libraries are derived from mRNA, which means they Show more…
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Imagine you want to study human crystallins, proteins present in the lens of the eye. To obtain a sufficient amount of the protein, you decide to clone the crystallin gene. Would you construct a genomic library or a cDNA library? What material would you use as a source of DNA or RNA?
Adi S.
Consider three different kinds of human libraries: a genomic library, a brain cDNA library, and a liver cDNA library. a. Suppose that all three of these libraries are sufficiently large so as to represent all of the different human nucleotide sequences that the library could possibly include. Which of these libraries would then correspond to the largest fraction of the total human genome? b. Would you expect any of these libraries not to overlap the others at all in terms of the sequences it contains? Explain. c. How do these three libraries differ in terms of the starting material for constructing the clones in the library? d. Why would you need to sequence many clones from many cDNA libraries to annotate a genome?
a) How does the construction of a genomic DNA library differ from that of a cDNA library? b) Give one advantage and one disadvantage of having constructed a mouse genomic library (rather than a cDNA library). c) You have constructed the mouse genomic library by cloning into the BamHI site of pUC19, and transformation into E. coli. Explain how you would use a combination of antibiotic and colour selection to identify colonies containing potential genomic clones or recombinant plasmids. d) You want to select a clone from your genomic library that contains gene A. It is known that gene A encodes protein A, with .... His – Cys – Met – Ser – Arg – Phe ..... as part of its amino acid sequence. You wish to make a set of 18 bp DNA probes to screen your DNA library with. i) Considering the degeneracy of the genetic code, how many possible different DNA sequences could encode this amino acid sequence? ii) Give the DNA sequence of any one probe that would be suitable to use. iii) You find that this probe you have designed does not detect any colonies in the genomic library, however it successfully binds to colonies from a cDNA library prepared from the same mouse. Give a possible explanation for this observation.
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