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TEM-1型β-内酰胺酶基因的克隆、表达及酶的纯化

Cloning, expression of a gene encoding TEM-1 P-lactamase and its purification

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【作者】 孙伟龚家玮朱春宝朱宝泉

【Author】 SUN Wei, GONG Jia-Wei, ZHU Chun-Bao, ZHU Bao-Quan(Shanghai Institute of Pharmaceutical Industry, Shanghai 200040)

【机构】 上海医药工业研究院上海医药工业研究院 上海 200040上海 200040上海 200040

【摘要】 开发专一性β-内酰胺酶抑制剂,对解决细菌耐药性问题具有重要意义。这项研究需要用到纯度较高的β-内酰胺酶。我们将TEM-1型β-内酰胺酶基因克隆到一个高表达载体中,研究了该酶在大肠杆菌中的高表达条件及分离纯化方法,最终获得的酶纯度大于90%。

【Abstract】 The development of new specific P-lactamase inhibitor is very important for overcoming the serious problem of bacterial resistance to p-lactam antibiotics used in clinic. High homogenous P-lactamase is needed in the process for screening small peptides which could bind to the active center of P-lactamase in vitro. The gene fragment encoding P-lactamase was cloned into a high expressing vector and P-lactamase was expressed in the form of inclusion body. Then the processes for wash, denaturation, refolding and purification of inclusion body were studied to obtain purified P-lactamase of more than 90% homogeneity.

【关键词】 β-内酰胺酶包涵体表达纯化
【Key words】 P-lactamaseinclusion bodyexpressionpurification
【基金】 国家自然科学基金(30171120)
  • 【文献出处】 中国医药工业杂志 ,Chinese Journal of Pharmaceuticals , 编辑部邮箱 ,2004年01期
  • 【分类号】Q78
  • 【被引频次】3
  • 【下载频次】152
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