Sonnewald U (1993)
Publication Status: Published
Publication Type: Journal article
Publication year: 1993
Publisher: SPRINGER VERLAG
Book Volume: 189
Pages Range: 174-181
Journal Issue: 2
Sucrose-phosphate synthase (SPS) from leaves of spinach (Spinacia oleracea L.) has been purified to homogeneity by a procedure involving precipitation with polyethylenglycol and chromatography over diethylaminoethylcellulose, OMEGA-aminohexylagarose, Mono Q and Blue Affinity columns. The purification factor was 838 and the final specific activity was 1.3 nkat . (mg protein)-1. On denaturing gels the major polypeptide was 120 kDa but there was also a variable amount of smaller polypeptides in the range of 90 to 110 kDa. A new activity stain was developed to allow visualization of SPS in gels. The holoenzyme had a molecular weight of about 240 and 480 kDa in native gels and Sepharose, respectively. A high-titre polyclonal antibody was obtained which reacted with SPS from other species including wheat, potato, banana and maize. Screening of a spinach-leaf cDNA-expression library with the antibody allowed the isolation of a full-length clone. Sequencing revealed a predicted molecular weight of 117649 Da, and considerable homology with the recently published sequence for maize leaf (Worrell et al. 1991, Plant Cell 3, 1121-1130). Expression of the spinach-leaf SPS gene in Escherichia coli resulted in biological activity, revealed by the presence of SPS activity in extracts and the accumulation of sucrose-6-phosphate and sucrose in the bacteria.
APA:
Sonnewald, U. (1993). PURIFICATION, CLONING AND EXPRESSION OF SPINACH LEAF SUCROSE-PHOSPHATE SYNTHASE IN ESCHERICHIA-COLI. Planta, 189(2), 174-181.
MLA:
Sonnewald, Uwe. "PURIFICATION, CLONING AND EXPRESSION OF SPINACH LEAF SUCROSE-PHOSPHATE SYNTHASE IN ESCHERICHIA-COLI." Planta 189.2 (1993): 174-181.
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