Below is a DNA fragment comprising the sequence of a gene coding for the human p

Question

Below is a DNA fragment comprising the sequence of a gene coding for the human protein CV H.

TTGATCGTACCTGCAGGTCAGAATTCCCAAGAAGCTTGGGCCCATATGTTTGCTGGGATCCATAGTGATTACAGTGGATCCTAGATTACTGCAGATAGCATGCATTAAGCTTAATTAGG

Write in the case the corresponding amino acid sequence of the protein (write in this manner Leu-Gly-Leu)  .

You would like to clone this gene in a plasmid next to a constitutive promoter (A constitutive promoter results in the constant transcription of a gene) and then insert this plasmid into bacteria in order to produce a high level of protein CV H. You have the choice between 3 plasmids: pA, pB and pC.

You also have the following enzymes:

EcoRI        G|AATTC

BamHI      G|GATCC

HindIII      A|AGCTT

PstI           CTGCA|G

Which plasmid would you use to clone your gene?

Which enzyme(s) would you use to cut the plasmid and the DNA fragment to facilitate the insertion of the fragment into the plasmid?

Promote Multiple Cloning Sites Sites are listed in order: . HindlII .BamHI .EcoRI .Pstl pA Promoter Multiple Cloning Sites Sites are listed in order: .Pstl .BamHI . HindlII .EcoRI Promoter Multiple Cloning Sites Sites are listed in order .EcoRI . Pstl . Hindlll .BamHI pc

Explanation / Answer

As the given sequence is a coding sequence therefore the

Coding Starnd is

5’ TTG ATC GTA CCT GCA GGT CAG AAT TCC CAA GAA GCT TGG GCC CAT ATG TTT GCT GGG ATC CAT AGT GAT TAC AGT GGA TCC TAG ATT ACT GCA GAT AGC ATG CAT TAA GCT TAA TTA GG 3’

mRNA

5’ UUG AUC GUA CCU GCA GGU CAG AAU UCC CAA GAA GCU UGG GCC CAU AUG UUU GCU GGG AUC CAU AGU GAU UAC AGU GGA UCC UAG AUU ACU GCA GAU AGC AUG CAU UAA GCU UAA UUA GG 3’

Amino acids

Leucine-isoleucine-valine-proline-alanine-glycine-glutamine-aspargine- serine-glutamine-glutamic acid- alanine-tryptophan-alanine-histidine-methionine-phenylalanine-alanine-glycine-isoleucine-histidine-Serine-aspartic acid-tyrosine-serine-glycine-serine- stop codon

There is stop codon encounters at this position, therefore, the the further synthesis of amino acid will not be occurred

Plasmid b (pB) will be used to clone the gene

The enzyme EcoRI will be used to cut the plasmid and the DNA fragment to facilitate the insertion of the fragment into the plasmid

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