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Fig. 1 | BMC Biotechnology

Fig. 1

From: A hyper-thermostable α-amylase from Pyrococcus furiosus accumulates in Nicotiana tabacum as functional aggregates

Fig. 1

PFA accumulates in Nicotiana tabacum in both transient expression and stable transformation. a Codon optimization improves accumulation of PFA in transient expression. Zymogram with tobacco leaf extracts transiently expressing pfa or an empty expression vector. Three replicates from three different plants are shown for each treatment. The bright bands indicate areas where starch is degraded by PFA. Each lane was loaded with 20 μl plant extract from 0.1 g leaf material extracted with 400 μl protein extraction buffer. PAGE-purified E.coli- produced PFA was used as standard. b Zymogram and (c) western blot hybridization analysis with anti-PFA antibodies of stable transgenic tobacco lines. PFA: 40 ng of E. coli-produced purified PFA. Lanes 1-8 represent eight independent transgenic lines from N. tabacum cv. TI95 (1-4 and 7-8) and I64 (5-6). The tobacco lines shown are representative of 16 N. tabacum cultivars used for producing 400 individual lines (25 lines/cultivar). WT: wild type, untransformed tobacco plant. Each lane was loaded with 5 μg total protein extracted with reducing extraction buffer

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