Recombination human ciliary neurotrophy factor, mutant and application thereof
A ciliary neurotrophic and mutant technology, which is applied in the fields of application, drug combination, and genetic engineering, can solve the problems of poor stability and low expression efficiency of expressed proteins, and achieve enhanced biological activity, increased stability, and easy purification and complexation. sexual effect
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[0018] The whole gene synthesis of the recombinant CNTF of the present invention is completed by Shanghai Sangon Bioengineering Technology Service Co., Ltd. When performing gene synthesis, it is required to design two enzyme cutting sites, NdeI and AvrII, respectively, at the upper and lower ends of the target gene. The synthesized target gene is 603bp in full length, loaded into the cloning vector PUC18 plasmid, the sequenced correct target gene is cut out, inserted into the expression vector pETr, and the protein is expressed and sequenced.
[0019] Compared with the natural human CNTF gene, the cDNA synthesized by the present invention replaces the cysteine at the 17th position of the human ciliary neurotrophic factor gene with alanine, and the glutamic acid at the 63rd position with arginine At the same time, part of the codons in the original CNTF gene were changed to codons preferred by prokaryotes, but no amino acid changes were involved, which was a synonymous mutatio...
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