CO差光谱法分析重组大肠埃希菌及产黄青霉中VHb的活性表达  

Quantitative analysis of Vitreoscilla hemoglobin expression in recombinant E.coli and Penicillium chrysogenum by carbon monoxide differential spectrum

作　　者：李兵[1] 周艳芬[1] 何叶喧[1] 赵晓瑜[1] 朱宝成[1] 

机构地区：[1]河北大学生命科学学院,保定071002

出　　处：《中国抗生素杂志》2005年第5期267-269,303,共4页Chinese Journal of Antibiotics

摘　　要：为快速、准确检测重组菌中透明颤菌血红蛋白(VHb)的活性表达,本文改进了CO差光谱法。诱导表达VHb的重组大肠埃希菌经超声破壁、高速离心和过量的Na2S2O4充分还原后,再在试样中通入CO与VHb进行络合反应,最后进行VHb的CO差光谱分析。VHb与CO络合物在419nm呈明显的吸收峰,因而可快速准确的检测VHb的活性表达。该方法亦适用于丝状真菌产黄青霉中VHb活性表达的检测。An improved carbon monoxide (CO) differential spectrometry assay method was established for the rapid determination of Vitreoscilla hemoglobin (VHb) expression. The recombinant E.coli which was induced to express VHb was sonicated to crash the cell, followed by ultra-centrifugation and then treated with sodium dithionite (Na2S2O4) , finally CO gas was bubbled to coordinate VHb and the differential spectrometry was analyzed. An absorption peak were observed. Expression result was determined rapidly and precisely. The method is also suitable for the determination of VHb expression of Penicillium chrysogenum.

关 键 词：透明产菌血红蛋白 CO差光谱 定量分析 产黄青霉 

分 类 号：Q344.13[生物学—遗传学]