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Figure 1 | BMC Immunology

Figure 1

From: Expression of human AID in yeast induces mutations in context similar to the context of somatic hypermutation at G-C pairs in immunoglobulin genes

Figure 1

Western blot analysis of hAIDSc expression in yeast. Yeast strain CG379-3-29RL transformed by expression vector pESC-LEU2 or pESC-LEU-hAIDSc were grown to logarithmic phase in a complete minimal medium without leucine. Then cells were washed and transferred into similar medium but containing galactose instead of glucose. Yeast protein extracts were prepared from approximately 200 mg of cells by the glass beads cell disruption method as described in [85]. Proteins were separated using 4–12% gradient PAA NuPage gel (Invitrogen). Transfer to PVDF membrane and reaction with primary antibodies (mouse anti c-myc) and then secondary antibodies (goat antimouse) was accomplished as suggested by the vendor (Western Breeze kit, Invitrogen). Lane 1 – Molecular weight markers (Benchmark, His-Tagged) were detected with antiHisx6 antibodies. Lane 2 – extract of yeast strain containing vector pESC-LEU. Lane 3 – extract of yeast strain containing pESC-LEU-hAIDSc.

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BMC Immunology

ISSN: 1471-2172

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