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Figure 5 | BMC Biotechnology

Figure 5

From: Efficient expression of codon-adapted affinity tagged super folder green fluorescent protein for synchronous protein localization and affinity purification studies in Tetrahymena thermophila

Figure 5

Western blot analysis of total proteins from T. thermophila cells expressing TtsfGFP, sfGFP, or EGFP. Equal amounts (30 μg) of EGFP (Lane 1), TtsfGFP (Lane 2), and sfGFP (Lane 3) were loaded. The left arrow shows a molecular mass of around 34 kDa corresponding to TtsfGFP. The quantity of TtsfGFP in the total protein extract appeared to be approximately 5–10-fold higher than sfGFP and EGFP. Total protein extracted from a Tetrahymena B2086 and CU428 cell mixture was used as a negative control (Lane 4). The positive control was TtsfGFP, which was constructed, expressed, and purified using Ni-NTA affinity purification from E. coli. The ~68 kDa band was predicted to be an sfGFP dimer (right arrow). Western blotting was performed with a monoclonal mouse anti-GFP antibody (1:1000). M: Bio-Rad Kaleidoscope western markers.

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BMC Biotechnology

ISSN: 1472-6750

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