Over Expression of Recombinant Staphylokinase and Reduction of Inclusion Bodies Using IPTG as Inducer in E. coli BL21 DE3

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Abstract

A previous research and studies have established the significance of describing Staphylokinase as a thrombolytic treatment, particularly in patients with cardiovascular disease, stroke, and other life-threatening disorders. Staphylokinase plays a significant part in the coagulation process through formation plasmin-Staphylokinase complex on the surface of the clot which activates plasminogen is generated by some strains of Staphylococcus aureus. The publication described the use of the E. coli strain BL12DE3 to produce the protein via the pET21a transporter is predicated on the use of a chemical stimulator (IPTG) which plays an important role in regulating protein recombinant by stimulating protein synthesis. Strain BL12DE3 activates T7 polymerase, encoding LacI via the pET21a transporter. It has been demonstrated that the IPTG inducer achieves the maximum level of protein expression in the shortest amount of time. A Staphylokinase was generated as a soluble protein in excess of 45 % by SDS-PAGE and subsequently purified by chromatography.
IPTG诱导重组葡萄激酶在大肠杆菌BL21 DE3中的过表达及包涵体的还原
先前的研究已经确定了将葡萄激酶描述为溶栓治疗的重要性,特别是在心血管疾病、中风和其他危及生命的疾病患者中。葡萄激酶通过在凝块表面形成纤溶酶-葡萄激酶复合物在凝块凝固过程中起重要作用,该复合物激活纤溶酶原是由某些金黄色葡萄球菌菌株产生的。该出版物描述了使用大肠杆菌菌株BL12DE3通过pET21a转运体产生蛋白质的前提是使用化学刺激物(IPTG),该刺激物通过刺激蛋白质合成在调节蛋白质重组中起重要作用。菌株BL12DE3激活T7聚合酶,通过pET21a转运体编码LacI。研究表明,IPTG诱导剂在最短的时间内实现了最高水平的蛋白表达。SDS-PAGE法得到可溶性蛋白含量超过45%的葡萄激酶,并用色谱法纯化。
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