TEM-1型β-内酰胺酶基因的克隆、表达及酶的纯化

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%.

Cloning, expression of a gene encoding TEM-1 β-actamase and its purification

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.