Question 9 6 pts Below is a DNA fragment comprising the sequence of a gene codin

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Question 9 6 pts Below is a DNA fragment comprising the sequence of a gene coding for the human protein CV H. ATTGCACGAATTCGTAGGTAGGGGATCCGTACGGTAGTTGAAGCTTAAGTCCACCATGTATCGAATTCCTTGATGTCAAGCTTTGTCTGCAGTTGTAAGTCCGGATCCTGCGTGGCATT 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. Promoter Muitiple Cloning Sites Sites are listed in order .BamHl .EcoRI .Pst pA Promoter Muitiple Cloning Sites Sites are listed in order .Pst .BamHl Hindl ECoRI Promoter Multple Cloning Sites Sites are listed in order .EcoRI pC Ps Hindl .BamH You also have the following enzymes: .ECORI GAATTC BamHI GGATCC HindilI AAGCTT PstlCTGCAI 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? You will be asked to explain your answer in the next question.

Explanation / Answer

5’-ATT GCA CGA ATT CGT AGG TAG GGG ATC CGT ACG GTA GTT GAA GCT TAA GTC CAC CAT GTA TCG AAT TCC TTG ATG TCA AGC TTT GTC TGC AGT TGT AAG TCC GGA TCC TGC GTG GCA TT-3’

5’-Isoleucine-Alanine-Arginine-Isoleucine- Arginine- Arginine-Stop Codon-Glycine- Isoleucine-Arginine-Thrionine-valine-valine-Glutamic-Alanine-stop codon-Valine-histidine-histidine-Valine-Serine-Asparagine-Serine-Leucine-Methionine-Serine-Serine-Phenylalanine-valine-Cysteine-Serine-Cysteine-Lysine-Serine-Glycine-Serine-Cysteine-Valine-Alanine-3’

Using Neb cutter tool we found the zero cutter enzyme suitable for this sequence which are given below:-

EcoR1        2 cutter

Bam H1     2 cutter

Hind3        2 cutter

Pst 1          1 cutter

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