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Influence of CpG content on gene expression.

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posted on 24.06.2020, 17:34 by Yusuke Ogaki, Miki Fukuma, Noriaki Shimizu

(A) CpG content of the sequences examined in this study. (B) Lambda-phage DNA was treated with bisulfite to convert unmethylated cytosine to uracil, followed by PCR amplification and cloning in E. coli host cells. The extent of conversion was determined by sequencing. (C) The repeat DNA of untreated or bisulfite-treated sequences was mixed with pΔBM.d2EGFP and co-transfected to CHO DG44 cells. (D) The B-3-31, untreated or bisulfite-treated lambda-phage sequence was cloned in pΔBM.d2EGFP and transfected to CHO DG44 cells. Cells were cultured in medium containing blasticidin for 24 (C) or 52 (D) days, and d2EGFP expression was analyzed using flow cytometry in the absence of butyrate. Unfilled lines indicate pΔBM.d2EGFP single transformants and gray-filled lines indicate the test transformants.