You discovered a novel eukaryotic organism that glows in the dark. You believe this trait is due to a single gene, and you wish to clone the gene. Which of the following strategies is most likely to be successful?
(A)Isolate the genomic DNA from the organism, digest with a restriction endonuclease, insert into a plasmid vector and transform into bacteria. Screen colonies for the ability to glow in the dark.
(B)Isolate the genomic DNA from the organism, digest with a restriction endonuclease, insert into a plasmid vector and transform into eukaryotic cells such as yeast. Screen colonies for the ability to glow in the dark.
(C)Isolate mRNA from the organism, reverse transcribe and generate cDNA, insert into a plasmid vector and transform into bacteria. Screen colonies for the ability to glow in the dark.
(D)Isolate mRNA from the organism, reverse transcribe and generate cDNA, insert into a plasmid vector and transform into eukaryotic cells such as yeast. Screen colonies for the ability to glow in the dark.Correct
Answer: (D)
Solution
Since this is a eukaryotic gene, its genomic copy likely contains introns that bacteria cannot splice out, so a cDNA (prepared from mature mRNA) must be
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