Assignment for STB1033

1. Discuss in brief a human genetic disease cause by mutation in the human DNA replication repair
system. (Hint: Go to Online Mendelian Inheritance in Man (OMIM) website for examples).

2. To do this question, you have to go the website below to retrieve the DNA sequence coding the
Homo sapiens phenylalanine hydroxylase (PAH) gene.

(http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=Retrieve&db=nucleotide&dopt=GenBank&list_uids=4557818)

Perform a web search for the restriction site that is recognized by the restriction enzyme NcoI.
Next, figure out how many NcoI sites are in the PAH sequence. In you answers, print out the whole
sequence and highlight the restriction sites. Also, mention the position of the cutting sites in your
answer.

3. A piece of DNA 900 bp long is cloned and then cut out of a vector for analysis. Digestion of this
linear piece of DNA with three different restriction enzymes singly and in all possible pairs of
enzymes gave the following restriction frgament size data. Construct a restriction map from these
data.
Enzymes Restriction Fragment Sizes
EcoRI
200 bp, 700 bp
HindIII
300 bp, 600 bp
BamHI
50 bp, 350 bp, 500 bp
EcoRI + HindIII
100 bp, 200 bp, 600 bp
EcoRI + BamHI
50 bp, 150 bp, 200 bp, 500 bp
HindIII + BamHI
50 bp, 100 bp, 250 bp, 500 bp