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Cell Biology: Nucelotide RNAase Digestion, Endonucleases

1. Digestion of the oligonucleotide, AGUACAA, by pancreatic RNase would yield what products?

2. A 5.5 kb, circular plasmid is cut with the following restriction endonucleases, producing the fragments listed. Construct a restriction map for this plasmid.

Endonuclease DNA fragments (kb)
Eco RI 5.5
Pst I 5.5
Bam HI 2.5 and 3.0
Eco RI and Bam HI 1.5 and 2.5
Eco RI and Pst I 0.5 and 5.0
Bam HI and Pst I 1.0, 2.0 and 2.5

Solution Preview

1. Digestion of the oligonucleotide, AGUACAA, by pancreatic RNase would yield what products?
<br>
<br>OTA: Pancreatic RNAase specifically cleaves after pyrimidines which are cytosine and uracil, so the products would be:
<br>
<br>AGU
<br>AC
<br>AA
<br>
<br>I have attatched a diagram of the mechanism of the cleavage. A brief description of the mechanism follows:
<br>
<br>RNAase has two essential His residues, His 12 and His 19, which act as acid and base catalysts. It is a two step process:
<br>
<br>1. His 12 takes a proton from the RNA 2'-OH group. The oxygen makes a nucleophilic attack on the ...

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