Preparation method of HPV16aE7/huhsp70 fusion protein
A technology for fusing proteins and proteins, applied in the fields of botanical equipment and methods, biochemical equipment and methods, DNA preparation, etc., can solve the problems of incorrect folding of foreign proteins, low yield of inclusion bodies, etc. good effect
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[0014] A preparation method of HPV16aE7 / huhsp70 fusion protein, comprising the following steps:
[0015] 1. Optimization and transformation of HPV16mE7 gene and gene cloning
[0016] Increase E7-specific MHC-I and MHC-II antigen epitopes on both sides of the E7 gene, pRb binding sites (Cys24Gly, Glu26Gly) in the E7 gene, and CKII phosphorylation sites related to transformation (Ser31Gly, Ser32Gly) ) and zinc finger binding region (Cys58Gly, Cys91Gly) mutations, the optimized and modified E7 gene (mE7) was obtained by gene synthesis, which can not only enhance the cellular immune and humoral immune response but also eliminate its transforming activity.
[0017] 2. Construction of pET30a-mE7, pET30a-huhsp70, pET30a-mE7 / huhsp70
[0018] The mE7 gene was synthesized by the gene synthesis method of Shanghai Jierui Biotechnology Co., Ltd., using the upstream primer P1 and downstream primer P2 of the mE7 gene, the mE7 gene was amplified by PCR and cloned into pMD18T, and then subclo...
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