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You are assigned the task to express an E. coli gene in cultured human cells. From published research, you know the amino acid sequence of the E. coli protein, which requires no post-translational modification.

2A. How would you modify the coding sequence of the E. coli gene in order to optimize its expression in human cells? This operation is done on a computer, not resulting in actual DNA molecules, but rather a sequence of nucleotides.

The coding sequence of the E. Coli gene must be modified with a Kozak sequence at 5 prime end because it aids in the initiation of translation.

2B. How will you actually produce the optimized DNA sequence in order to clone it in an expression vector for human cells? Describe the methods in conceptual terms. Use as simple a method as possible. Do not skip important steps.

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Tod Thiel
Tod ThielLv2
28 Sep 2019

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