You are trying to find the gene responsible for a human genetic disorder. You ha

Question

You are trying to find the gene responsible for a human genetic disorder. You have mapped the gene to a particular chromosome region, and examining the human genome sequence for that region gives you the
nucleotide sequence below:
5’ CATACTTACTACTAGATTACGATTAGACGATTAGGATGGCCGACTCGTGCAGTAAC
AGCATGACCGAGGCCTAGACCAGATTAGGAGCCGGACCAGGACGGACCAGCGACT 3’
a. Assuming you are reading the non-coding strand and that there are no introns, find an open reading frame (ORF) in this region. Circle the point where translation will start, and put a box around the point where translation will stop. Then give the number of amino acids in the protein this gene would encode:

b. If you wanted to express this gene in E. coli, what would need to be present in your cloning vector to ensure that it will be transcribed and translated?

c. How might the protein produced by E. coli differ from the protein produced from the same gene in a
human cell?

Explanation / Answer

a). Template strand-

5’ CATACTTACTACTAGATTACGATTAGACGATTAGGATGGCCGACTCGTGCAGTAAC
AGCATGACCGAGGCCTAGACCAGATTAGGAGCCGGACCAGGACGGACCAGCGACT 3’

open reading frame -

5’ G U A U G A A U GAUG A U C U A A U G C U A A UC UG C U A A U C C U A C C G G C U G A G C A C G U C U T T G U C G U A C U GGC U C C G G A U C U G G U C U AA U C C U C G G C C U G G U C C U G C C U G G U C G C U G A 3’

Translation starts at AUG and ends when a non-coding strand is encountered.

5’ G U A U G A A U G AUG A U C U A A U G C U A A UC UG C U A A U C C U A C C G G C U G A G C A C G U C U T T G U C G U A C U GGC U C C G G A U C U G G U C U AA U C C U C G G C C U G G U C C U G C C U G G U C G C U G A 3’

AUG - initiates the translation process.

UAA - terminates the translation.

Amino acids that are coded are -

AUG - methionine

AUC- leucine

b). To express this gene in E.coli the eukaryotic coding sequence has to be placed after prokaryotic promoter and ribosome building site.

c). The protein produced by E. coli differ from the protein produced from the same gene in a
human cell in its post translation modifications. They are different in both. In eukaryotes more translation modifications occurs than in prokaryotes.

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