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- 2025年07月24日
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文献和实验Identification of Asp Isomerization in Proteins by 18O Labeling and Tandem Mass Spectrometry
to identify the site of modification by LC-MS/MS peptide mapping. Here, we describe an approach to label the Asp residue involved in isomerization at the protein level by hydrolyzing the succinimide intermediate in H 2 18 O. Tryptic digestion of this labeled
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
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