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

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. . pmid:1697508. PubMed HubMed [Campbell-1990]

Notes

Recent changes