Islet-targeted protein and eukaryotic expression method thereof
An expression method and protein technology, which are applied in the field of pancreatic islet-targeted protein and eukaryotic expression of pancreatic islet-targeted protein, can solve the problem of preparing soluble IGRP206-214MHC complex, prokaryotic expression system without glycosylation function, and unable to truly simulate protein features, etc.
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[0029] Preferred embodiments of the present invention will be described in more detail below. Wherein, various reagents used can be obtained commercially, and the specific operation method is carried out with reference to the operation manual attached to the commercially available reagents.
[0030] The term abbreviations, English full names and Chinese full names involved in the present invention are shown in Table 1.
[0031] Table 1
[0032]
[0033] 1. IGRP 206-214 - Construction of dtSCT eukaryotic expression system and protein preparation
[0034] amplified IGRP 206-214 The full-length coding gene, adding a biotinylated site and 6×his tag at the end, and adding a restriction site at the beginning and end, inserted into the eukaryotic expression plasmid, and introduced into Pasteur yeast to construct a eukaryotic expression system for the protein, And use methanol culture to induce the expression of the target protein.
[0035] Use Oligo 7 to design the upstream a...
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