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Question 2. You want your recombinant Human Growth Hormone to be tagged at the N-terminus with a poly- histidine domain (which is present in the vector), so you need your to cloned in a way that maintains the reading frame. To facilitate cloning, you have added an EcoR1 restriction site to the N-terminus of the Human growth hormone, as an EcoR1 site is also present in the vector (EcoR1 sites in red; N-terminus of HGH in blue). Vector 1: Vector 2: Vector 3: i) Answer: ii) His His His His His His Gly Asn Ser Gly Gln CAT CAT CAT CAT CAT CAT GGG AAT TCG GGC CAA DNA to be inserted into the vector: Answer: His His His His His His Glu Phe Gly Pro Lys CAT CAT CAT CAT CAT CAT GAA TTC GGG CCC AAA His His His His His His Gly Ile Arg Ala Gln CAT CAT CAT CAT CAT CAT GGA ATT CGG GCC CAA Which vector will you use, 1, 2, or 3? Met Ala Thr Gly... AACAAGAATTCTATG GCT ACA GGG... Draw the final DNA (bottom) and tagged protein (top) sequences after you have ligated the insert to the proper vector

Question 2. You want your recombinant Human Growth Hormone to be tagged at the N-terminus with a poly- histidine domain (which is present in the vector), so you need your to cloned in a way that maintains the reading frame. To facilitate cloning, you have added an EcoR1 restriction site to the N-terminus of the Human growth hormone, as an EcoR1 site is also present in the vector (EcoR1 sites in red; N-terminus of HGH in blue). Vector 1: Vector 2: Vector 3: i) Answer: ii) His His His His His His Gly Asn Ser Gly Gln CAT CAT CAT CAT CAT CAT GGG AAT TCG GGC CAA DNA to be inserted into the vector: Answer: His His His His His His Glu Phe Gly Pro Lys CAT CAT CAT CAT CAT CAT GAA TTC GGG CCC AAA His His His His His His Gly Ile Arg Ala Gln CAT CAT CAT CAT CAT CAT GGA ATT CGG GCC CAA Which vector will you use, 1, 2, or 3? Met Ala Thr Gly... AACAAGAATTCTATG GCT ACA GGG... Draw the final DNA (bottom) and tagged protein (top) sequences after you have ligated the insert to the proper vector

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
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Question 2. You want your recombinant Human Growth Hormone to be tagged at the N-terminus with a poly- histidine domain (which is present in the vector), so you need your to cloned in a way that maintains the reading frame. To facilitate cloning, you have added an EcoR1 restriction site to the N-terminus of the Human growth hormone, as an EcoR1 site is also present in the vector (EcoR1 sites in red; N-terminus of HGH in blue). Vector 1: Vector 2: Vector 3: i) Answer: ii) His His His His His His Gly Asn Ser Gly Gln CAT CAT CAT CAT CAT CAT GGG AAT TCG GGC CAA DNA to be inserted into the vector: Answer: His His His His His His Glu Phe Gly Pro Lys CAT CAT CAT CAT CAT CAT GAA TTC GGG CCC AAA His His His His His His Gly Ile Arg Ala Gln CAT CAT CAT CAT CAT CAT GGA ATT CGG GCC CAA Which vector will you use, 1, 2, or 3? Met Ala Thr Gly... AACAAGAATTCTATG GCT ACA GGG... Draw the final DNA (bottom) and tagged protein (top) sequences after you have ligated the insert to the proper vector
Transcribed Image Text:Question 2. You want your recombinant Human Growth Hormone to be tagged at the N-terminus with a poly- histidine domain (which is present in the vector), so you need your to cloned in a way that maintains the reading frame. To facilitate cloning, you have added an EcoR1 restriction site to the N-terminus of the Human growth hormone, as an EcoR1 site is also present in the vector (EcoR1 sites in red; N-terminus of HGH in blue). Vector 1: Vector 2: Vector 3: i) Answer: ii) His His His His His His Gly Asn Ser Gly Gln CAT CAT CAT CAT CAT CAT GGG AAT TCG GGC CAA DNA to be inserted into the vector: Answer: His His His His His His Glu Phe Gly Pro Lys CAT CAT CAT CAT CAT CAT GAA TTC GGG CCC AAA His His His His His His Gly Ile Arg Ala Gln CAT CAT CAT CAT CAT CAT GGA ATT CGG GCC CAA Which vector will you use, 1, 2, or 3? Met Ala Thr Gly... AACAAGAATTCTATG GCT ACA GGG... Draw the final DNA (bottom) and tagged protein (top) sequences after you have ligated the insert to the proper vector
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