User:Torsten Waldminghaus/Notebook/Chip on Chip with SeqA

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Project Description/Abstract

SeqA is a DNA binding protein that binds specifically to hemimethylated GATC sites in E. coli and other γ-proteobacteria. Analysis of the chromosome wide binding in vivo could provide further information on its function beyond regulation of replication initiation. For Chip on chip experiments (see ChIP-Chip E. coli) one only needs a good antibody and an array. In the first experiments the SeqA ab turned out to also detect RNA-polymerase or some associated protein. This means that good purification is needed before repeating the experiments. One try is an affinity substraction Affinity purification of antibodies.

Future Plans

  • Compare SeqA binding in fast and slow growing cells
  • Analyze SeqA binding in synchronized cells, if possible with the Baby cell column.

References

  1. Campbell JL and Kleckner N. E. coli oriC and the dnaA gene promoter are sequestered from dam methyltransferase following the passage of the chromosomal replication fork. Cell. 1990 Sep 7;62(5):967-79. PubMed ID:1697508 | HubMed [Campbell-1990]

Notes

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