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. 2017 Sep 22;18(10):2035. doi: 10.3390/ijms18102035

Figure A1.

Figure A1

The expression, purification and identification of recombinant lamprey Vav3 his-tag fusion protein. A pET-32a (+) vector was used as the expression vector for expressing lamprey Vav3 open reading frame (ORF) region in Escherichia coli BL21 (DE3) as a fusion protein with 6× His-tag. The recombinant lamprey Vav3 was separated by 12% SDS-PAGE and stained with coomassie brilliant blue R-250. (a) The recombinant lamprey Vav3 was overexpressed in E. coli BL21 (DE3) strain by 1 and 0.1 mM IPTG induction. M: protein marker; 1: crude lysate of E. Coli BL21 (DE3) transformed by pET-32a (+) vector with 1mM IPTG induction; 2: crude lysate of E. Coli BL21 transformed by pET-32a (+lamprey Vav3) vector without IPTG induction; 3: crude lysate of E. Coli BL21 transformed by pET-32a (+lamprey Vav3) vector with 1mM IPTG induction; 4: crude lysate of E. Coli BL21 transformed by pET-32a (+lamprey Vav3) vector with 0.1mM IPTG induction; (b) The recombinant lamprey Vav3 his-tag fusion protein was purified by using affinity chromatography method with Ni-NTA His-Bind® Resin. M: protein marker; 1: target protein eluted with 25 mM imidazole elution buffer; 2: target protein eluted by 100 mM imidazole elution buffer; (c,d) Peptide mass fingerprinting analysis of recombinant lamprey Vav3 protein bands 1 and 2 by an autoflex™ speed MALDI-TOF mass spectrometer.