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. 2001 Apr;12(4):901–917. doi: 10.1091/mbc.12.4.901

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Copyright © 2001, The American Society for Cell Biology

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Detection of Spo20 and Spo20^G275D by Western blotting. (A) Strategy for constructing the spo20-HA and spo20^G275D-HA fusion genes. (B) Western blotting analysis of Spo20 and Spo20^G275D. Wild-type strain (YN8-WH) and spo20 mutant strain (YN8-MH) carrying spo20-HA and spo20^G275D-HA, respectively, were cultured in liquid sporulation medium (MM-N) at 25°C. Protein extracts were prepared at 0 and 10 h (lanes 1–4). Cells were also cultured in liquid complete medium (YEL) at 25 or 37°C for 12 h (lanes 5–8). Protein extracts were subjected to immunoblot analysis with the mouse anti-HA, as well as with anti-α-tubulin antibody as the loading control. Lanes 1, 3, 5, and 7, Spo20-HA; lanes 2, 4, 6, and 8, Spo20^G275D-HA. Spo20-HA was detected as a band of ∼39 kDa.