Advisor(s) - Committee Chair
Valgene Dunham, Frank Toman, Martin Houston
Department of Biology
Master of Science
Two forms of glutamine synthetase (GS) have been isolated from three-day old etiolated soybean hypocotyls by ammonium sulfate precipitation and DEAE-cellulose chromatography. The GS isoforms were eluted from a 30 ml DEAE-cellulose column using 0.14 M KC1 and 0.175 M KC1 and from a 1.0 ml DEAE-cellulose column using 0.08 M KC1 and 0.14 M KC1. The two isoforms of soybean GS have a molecular weight of about 392,000 daltons as indicated by in gel enzyme assays and staining of protein bands.
The poly(A)+RNA specific for GS synthesis was isolated using phenol and chloroform followed by Hybond mAP paper. Translation was performed using a wheatgerm cell-free lysate. Following the application of the in vitro translated products to a 1 ml DEAE-cellulose column, GS was successfully eluted at 0.14 M KC1.
Biology | Life Sciences
Silver, Stanley, "Glutamine Synthetase: Isolation of Isoforms, Poly (A) +RNA and In Vitro Translation" (1987). Masters Theses & Specialist Projects. Paper 2852.