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Interconversion of intra‐ and extra‐chromosomal sites of gene amplification by modulation of gene expression and DNA methylation by Noriaki Shimizu; Naoyuki Hanada; Kohichi Utani; Naoki Sekiguchi is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Interconversion of intra‐ and extra‐chromosomal sites of gene amplification by modulation of gene expression and DNA methylation about?

## Abstract We previously showed that plasmids containing a mammalian replication initiation region and a matrix attachment region were efficiently amplified to few thousand copies per cell, and that they formed extrachromosomal double minutes (DMs) or chromosomal homogeneously staining regions (HSRs). In these structures, the plasmid sequence was arranged as a tandem repeats, and we suggested a mechanism of plasmid amplification. Since amplification was very efficient, easy, and convenient, it might be adapted to a novel method for protein production. In the current study, we found that gene expression from the tandem plasmid repeat was suppressed. We identified several strategies to overcome this suppression, including: (1) use of higher concentrations of antibiotic during cell selection; (2) treatment of cells with agents that influence DNA methylation (5‐azacytidine) or histone acetylation (butyrate); (3) co‐amplification of an insulator sequence; and (4) co‐amplification of sequences that encode a transcriptional activator. Expression from the plasmid repeat was always higher at DMs compared to HSRs. We found that continuous activation of a plasmid‐encoded inducible promoter p

Who reads Interconversion of intra‐ and extra‐chromosomal sites of gene amplification by modulation of gene expression and DNA methylation?

Author

Noriaki Shimizu; Naoyuki Hanada; Kohichi Utani; Naoki Sekiguchi

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; Wiley (ISSN 0730-2312)

Published

2007

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)