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Integrating cloning and expression

PGR amplification of a gene from prokaryotic DNA is best achieved by prior isolation of genomic DNA from the prokaryote of interest in the following way. In this case the sodB [Pg.168]

Successful transformation with a recombinant pDNA expression construct should result in the appearance of a number of colonies on the antibiotic selection plate, with none appearing [Pg.169]

For higher eukaryotes, the most common approach to cloning is PCR amplification from cDNA libraries in order to avoid the intron problem. Another challenge when expressing eukaryotic proteins is that eukaryotic proteins frequently do not fold properly in E. coli and will not be processed with the correct post-translational modifications, and therefore often other eukaryotic expression hosts must be used. Options include using yeast cells, insect cell lines or mammalian cell lines. [Pg.170]


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