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大鼠SM22α的原核表达与纯化

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摘要 目的构建大鼠平滑肌22α(smooth muscle 22α,SM22α)原核表达质粒,使其在原核细胞中高效表达并纯化。方法将SM22αcDNA全长片段插入pGEX-3X载体构建重组子pGEX3X-SM22α,转化宿主菌。经异丙基硫代--βD-半乳糖苷诱导表达SM22α蛋白,谷胱甘肽-S-转移酶(glutathione-S-transferase,GST)亲和层析纯化。结果重组子经酶切后,电泳分离出预期长度的插入片段。重组子转化的大肠杆菌JM109,经诱导剂诱导和GST亲和层析纯化后,得到纯化的GST-SM22α融合蛋白。结论本实验重组的pGEX3X-SM22α可在大肠杆菌中高效表达GST-SM22α融合蛋白,为进一步研究SM22α的功能奠定了基础。
出处 《河北医科大学学报》 CAS 2007年第5期354-355,共2页 Journal of Hebei Medical University
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参考文献3

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