组氨酸标签位置对重组鹰嘴豆孢克鲁维酵母外切菊粉酶活性的影响

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Publication date
1 January 2016
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Abstract

为了研究His_6-tag标签位置对鹰嘴豆孢克鲁维酵母CBS4857外切菊粉酶(Kluyveromyces cicerisporus CBS4857 exo-inulinase,KcINU1)酶活性的影响,实验首先根据Swiss-Model构建的KcINU1三维模型预测了组氨酸标签在KcINU1的位置,然后将编码His_6-tag的基因通过PCR方法分别引入到kcINU1的N端和C端,并将重组的kcINU1基因克隆到毕赤酵母表达载体pPICZ&alpha;A,重组质粒进一步经电转化法整合到毕赤酵母宿主菌X-33中,最后用甲醇诱导进行分泌表达,镍柱亲和层析法纯化目的蛋白,DNS去进行外切菊粉酶的活性测定。结...</p

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