Preparation of heteroduplex enhanced green fluorescent protein plasmid for in vivo mismatch repair activity assay

Bisheng Zhou, Changjiang Huang, Junhua Yang, Jianxin Lu, Qiaoxiang Dong, Lu Zhe Sun

Research output: Contribution to journalArticle

22 Scopus citations

Abstract

Preparation of heteroduplexes in large quantities with high purity is essential for the measurement of DNA mismatch repair (MMR) activity. Here we report a rapid, less labor-intensive method for the preparation of a heteroduplex plasmid that expresses the enhanced green fluorescent protein (EGFP) if the mismatch is repaired correctly. The method involves the use of a wild-type and a mutated EGFP expression plasmid and a few steps of enzymatic digestion. When the constructed heteroduplex EGFP plasmid was transfected into MMR-proficient and -deficient cell lines, the number of EGFP-expressing cells was much higher in the MMR-proficient cells than in the MMR-deficient cells, suggesting that the heteroduplex can be used for MMR activity assay in live model systems.

Original languageEnglish (US)
Pages (from-to)167-169
Number of pages3
JournalAnalytical Biochemistry
Volume388
Issue number1
DOIs
StatePublished - May 1 2009

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ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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