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利用 L-ans B基因片段构建了融合表达载体 p EC,其中 L-ans B基因中编码唯一酸水解位点的序列已通过定点突变消除 ,并在它的 3′端引入了新的酸水解位点编码序列和方便外源基因插入的克隆位点 Bam H 和 Hind ,外源基因可从克隆位点 Bam H 和 Hind 处插入 p EC中 ,表达的融合蛋白用酸水解法加工后仅被切割成一大一小两个片段 ,不会导致产物混杂。 h GRF对它进行验证的结果显示 h GRF基因不仅能按正确的读码框插入 ,而且表达的融合蛋白与预期大小一致 ,说明 p EC用于融合表达外源肽基因是有效的可行的
The fusion expression vector p EC was constructed by using the L-ans B gene fragment, in which the sequence encoding the sole acid hydrolysis site in L-ans B gene was eliminated by site-directed mutagenesis and a new acid hydrolysis site was introduced at its 3 ’end Point coding sequence and facilitate the insertion of exogenous gene cloning sites Bam H and Hind exogenous genes can be inserted from the cloning site Bam H and Hind at p EC in the expression of the fusion protein after acid hydrolysis process was only cut into a Freshman two small pieces, will not lead to product confusion. h GRF showed that the h GRF gene can not only be inserted into the correct reading frame, but also the expressed fusion protein with the expected size, indicating that p EC is effective and feasible for the fusion expression of the foreign peptide gene