protein mbp fusion expression vector pmal c2  (New England Biolabs)

Journal: Neural Regeneration Research

Article Title: Two memory associated genes regulated by amyloid precursor protein intracellular domain

doi: 10.3969/j.issn.1673-5374.2012.05.003

Figure Lengend Snippet: The maltose-binding protein (MBP)-amyloid precursor protein intracellular domain (AICD) fusion protein was expressed, purified and detected in vitro . (A) Whole-cell extracts from E. coli BL21 transformed with AICD/pMAL-c2 plasmids either noninduced (lane I) or induced by isopropyl-β-D-thiogalactoside (lane II), purified MBP-AICD (lane III) and MBP (lane IV) were separated on 10% sodium dodecyl sulfate-polyacrylamide electrophoresis gels and stained with Coomassie brilliant blue. (B) MBP-AICD (lane I, corresponding to lane III in ) but not MBP (lane II, corresponding to lane IV in ) was recognized by a specific antibody against AICD in western blot analysis.

Article Snippet: The DNA fragment encoding AICD59, modified by the addition of a Bam HI (New England Biolabs (Beijing) Ltd., Beijing, China) site at the 5′ terminus and a Hind III (New England Biolabs (Beijing) Ltd.) site at the 3′ terminus, was amplified via PCR from full-length APP695 (New England Biolabs (Beijing) Ltd.) and then cloned into the MBP fusion expression vector pMAL-c2 (New England Biolabs (Beijing) Ltd.).

Techniques: Binding Assay, Purification, In Vitro, Transformation Assay, Electrophoresis, Staining, Western Blot