Production of Abrus pulchellus ribosome-inactivating protein from seeds callus culture

摘要: Ribosome inactivating proteins (RIPs) were isolated from callus culture that established seed explants of Abrus pulchellus. Cotyledon segments immature seeds inoculated in basal medium MS supplemented with different concentrations auxin (2,4-D), citokinin (kinetin and BA) sucrose order to determine the best induction. A. pulchellus type 2 RIP (pulchellin) expression was monitored cultures by RT-PCR biological activity. The calli obtained after 35 days freeze dried, macerated submitted extraction total RNA (0.1 M Tris-HCl pH 7.6 buffer, containing 0.15 NaCl, 3 h at room temperature). A specific DNA fragment codifying A-chain pulchellin amplified suggesting presence protein. This confirmed crude extract showed haemagglutinating activity against rabbit blood cells a high intraperitoneal toxicity mice. also affinity chromatography on Sepharose-4B column. retained protein, peak released 0.1 galactose, appeared be composed two main bands polyacrylamide gel electrophoresis, denaturating conditions, similar pattern seeds.