1. Companion site for Biotechnology. by Clark Copyright © 2009 by Academic Press. All rights reserved. 1 Expression of Eukaryotic Proteins A bacterial Promoter/terminator must be used Bacteria can’t process introns, so cDNA must be used Translational Expression Vector Insertion of your gene into the MCS must be done just right to get the RBS at the optimum 8 bases away from start ATG Strong means that sometimes multiple termination sequences are used

2. Companion site for Biotechnology. by Clark Copyright © 2009 by Academic Press. All rights reserved. 2 Codon Usage Affects Rate of Translation. Bacteria prefer one codon for a particular amino acid to other redundant codons. In this example, the ribosome is stalled because it is waiting for lysine tRNA with a UUU anticodon. FIGURE 10.3

3. Companion site for Biotechnology. by Clark Copyright © 2009 by Academic Press. All rights reserved. 3 http://bio.takara.co.jp/product/basic_info.asp?catcd=B1000420&subcatcd=B1000421&unitid=U100006426

4. Companion site for Biotechnology. by Clark Copyright © 2009 by Academic Press. All rights reserved. 4

5. Companion site for Biotechnology. by Clark Copyright © 2009 by Academic Press. All rights reserved. 5 Comparison of Recombinant Protein Expression SystemsEach protein expression system falls on a continuum of worst to best for characteristics such as speed, cost, glycosylation, folding, and government regulations. Transgenic animals (rabbit) and transgenic plants (plants) are discussed in Chapters 14 and 15 chapter 14 chapter 15. The other symbols include mammalian cultured cells, insect cell culture, yeast, and bacteria. FIGURE 10.15