Fusion tagging protein for nonchromatographic sepration of target protein and coding gene and preparation method thereof
A chromatographic separation and fusion tag technology, which is applied in the fields of fusion tag proteins and their encoded genes and preparation, can solve the problems of pollution, mixing, and long reaction time, and achieve the effects of avoiding cumbersome operations, high-efficiency expression, and low cost.
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[0023] A fusion tag protein capable of non-chromatographic separation of target proteins, characterized in that it has the amino acid sequence shown in Sequence Table 1 or its fusion protein, or its functional fragment, or one of its functional fragment derivatives kind. A preparation method of a fusion-tagged protein (KV8F) capable of performing non-chromatographic separation of the target protein in this embodiment: the coding gene of the sequence shown in Sequence Table 2 is inserted into the modified high-efficiency expression vector pET-22b. The transformation method is to use the gene fragment CAT ATG AGC AAA GGG CCG GGC TGG CCG TGA TAA GAA TTC in the sequence table 3 to replace the gene fragment from Nde I to EcoR I in pET-22b (+). Restriction digestion with pf1MI and Bg1 I, recovery of ELP[KV after electrophoresis separation 8 F-20] gene fragment, and then introduce this gene into the pET-22b (+) expression vector modified with SfiI enzyme digestion to construct the p...
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