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发明公开
CN109810983A 猪INF-γ的优化基因和采用该优化基因制备猪INF-γ的方法及其注射液和应用 有权

猪INF-γ的优化基因和采用该优化基因制备猪INF-γ的方法及其注射液和应用
摘要：
本发明公开了一种猪INF-γ的优化基因和采用该优化基因制备猪INF-γ的方法及其注射液和应用，所述猪INF-γ的优化基因的基因序列如SEQ ID NO.1所示，所述制备方法包括：1)将权利要求1所述的基因序列克隆到真核表达载体中，得到重组真核表达载体；2)将步骤1)中所述重组真核表达载体转染CHO细胞，通过培养、筛选和驯化，得到高效表达猪INF-γ的CHO细胞株；以及3)将步骤2)中所述CHO细胞株进行发酵培养，从发酵培养上清中纯化得到猪INF-γ。所述注射液包括60μg猪INF-γ与70佐剂。使用优化后的基因序列在CHO细胞中制备猪INF-γ蛋白时具有产量高、易纯化、成本低、质控容易，且具有良好的活性，可以作为免疫增强剂和治疗药品使用。
摘要（英）：

The invention discloses an optimized gene of pig INF-gamma and a method for preparing the pig INF-gamma by using the optimized gene as well as injection and application of the optimized gene. The genesequence of the optimized gene of the pig INF-gamma is shown as SEQ ID NO. 1. The preparation method comprises the following steps: firstly, cloning the gene sequence in claim 1 into an eukaryotic expression vector to obtain a recombinant eukaryotic expression vector; secondly, transfecting CHO cells by the recombinant eukaryotic expression vector in the first step, and culturing, screening and domesticating to obtain a CHO cell strain for efficiently expressing the pig INF-gamma; thirdly, carrying out fermentation culture on the CHO cell strain in the second step and purifying fermentation culture supernatant to obtain the pig INF-gamma. The injection is prepared from 60 mu mg pig INF-gamma and Merckinade 70 adjuvant. By using the optimized gene sequence to prepare the pig INF-gamma protein in CHO cells, the method has the advantages of high yield, easiness in purification, low cost and easiness in quality control; in addition, the pig INF-gamma injection has good activity, and can be used as an immunoenhancer and a therapeutic drug.

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