Ghosh Swati, Thakur Mahendra Kumar
Biochemistry and Molecular Biology Laboratory, Center of Advanced Study in Zoology, Banaras Hindu University, Varanasi, 221005, India.
Mol Biol Rep. 2008 Dec;35(4):589-94. doi: 10.1007/s11033-007-9128-9. Epub 2007 Sep 3.
Escherichia coli (E. coli) is the most widely used prokaryotic host system for the synthesis of recombinant proteins. The overproduction of recombinant proteins is sometimes lethal to the host cells. In the present study, we expressed the ligand binding domain (LBD) of mouse estrogen receptor alpha (mouse ERalpha) using an expression vector (pIVEX) in E. coli BL21(DE3) and examined the effect of production of this protein on bacterial growth. The expressed protein was immunologically detected as a 30 kD histidine-tagged protein in the soluble part of the bacterial lysate. The overproduction of mouse ERalpha-LBD, as reflected by total protein content and expression pattern, resulted in the decrease of bacterial growth.
大肠杆菌(E. coli)是用于合成重组蛋白的最广泛使用的原核宿主系统。重组蛋白的过量生产有时对宿主细胞是致命的。在本研究中,我们使用表达载体(pIVEX)在大肠杆菌BL21(DE3)中表达了小鼠雌激素受体α的配体结合域(小鼠ERα),并研究了该蛋白的产生对细菌生长的影响。表达的蛋白在细菌裂解物的可溶部分中被免疫检测为一种30 kD的组氨酸标签蛋白。从小鼠ERα-LBD的总蛋白含量和表达模式反映出的过量生产导致了细菌生长的下降。