Overexpression of mouse estrogen receptor-β decreases but its transactivation and ligand binding domains increase the growth characteristics of E. coli.
Escherichia coli is one of the most common and widely used prokaryotic hosts for the expression of recombinant proteins. The overexpression of recombinant proteins occasionally increases bacterial growth but sometimes reduces it and becomes lethal to the host cells. Here, we report the overexpression of mouse ER-β and its domains in the prokaryotic expression system and its opposite effect on the growth characteristics of E. coli. ER-β protein was immunologically detected as a 53 kDa his-tag protein in the pellet of the bacterial lysate. Its overexpression, as reflected by the total protein content and expression pattern, resulted in the decrease of bacterial growth. However, the overexpression of ER-β transactivation domain (TAD) using pIVEX and ligand binding domain (LBD) using pRSETA in E. coli BL21 (DE3) show opposite pattern. TAD was immunologically detected as 20 kDa and LBD as 22 kDa protein in the supernatant of the bacterial lysate and their overexpression increased the bacterial growth.
Paramanik
Vijay
V
Biochemistry and Molecular Biology Laboratory Centre of Advanced Study, Department of Zoology, Banaras Hindu University, Varanasi, 221005, India.
Thakur
M K
MK
D000818:Animals; D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development*; D047629:Estrogen Receptor beta / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D051379:Mice; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation
D047629:Estrogen Receptor beta; D011994:Recombinant Proteins