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coli containing pET-28b(+) based plasmids (pANT13-16, pANT18, pANT20-21, pANT35, pANT95, pJK1-3, pJK7-9 or site saturation variants of pANT35) were inoculated from an overnight preculture at a ratio of 1:100 in 96-deep-well plates equipped with a CR1296 sandwich cover (EnzyScreen) in 0.5 ml TB medium containing 50 g ml 1 kanamycin sulfate per well and incubated at 37 C, 300 rpm using a 5 cm shaking diameter for 2.5 h

These variants may facilitate the presentation of self-antigens (e.g., insulin peptides) to T cells and initiate an autoimmune cascade against pancreatic beta cells [2, 3]

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