Essay
A scientist wishes to produce a mammalian protein in E. coli. The protein is a glycoprotein with a molecular weight of 40,000. Approximately 20% of its mass is polysaccharide. The isolated protein is usually phosphorylated and contains three disulfide bonds. The cloned gene contains no introns. (a) What sequences or sites will be required in the vector to get this gene regulated, transcribed, and translated in E. coli? (b) List two problems that might arise in producing a protein identical to that isolated from mammalian cells and describe each problem in no more than two sentences.
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