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- 2025年07月10日
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文献和实验Recombinant DNA Engineering, Or Cloning Genes In Plasmids
and thus fusion proteins. For example there might be sequence for a fluorescent protein such as GFP or a peptide tag such as HA upstream of MCS (and downstream of promoter) so that an insert in frame will lead to the generation of a fusion protein, an N-terminal
, and usually final protein yield between 0.1 and 2 mg/g cells is acceptable. Another concern is protein degradation. Especially with Pichia , protease activity during cell lysis and purification is always an issue. The importance of N-terminal degradation
The Use of Recombinant Fusion Proteases in the Affinity Purification of Recombinant Proteins
of interest, but some affinity tails are able to be linked to either the N- or C-terminus of the protein of interest. The choice of affinity tail to use for the expression of any particular protein is empirical since the factors leading to the high expression of recombinant
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