Leitung: Prof. Dr. Thomas Noll
wiss. Mitarbeiter: Jennifer Becker, Oliver Rupp, Christina Timmermann
Normalized cDNA libraries originating from several CHO cell lines cultivated under different culture conditions were sequenced using the Roche 454 approach resulting in 1.84 million readswith an average read length of 373 nucleotides. The reads were assembled and grouped into 30,578 isotigs (transcripts) and 25,378 isogroups (genes). Taxonomic classification os the isotigs showed that 80% of the assembled data is most similar to the transcriptome of mouse. Coverage analysis demonstrated that 8,361 mouse transcripts, representing 6,193 genes, are covered by <95% of their sequence length by CHO isotigs.
Metabolic pathway reconstruction and analysis of N-glycosylation related biosynthesis routes revealed complete detection of all relevant genes. On the basis of the sequencing data a customized CHO cell microarray was designed. Probe functionality was shown using self-hybridization experiments. The customized CHO cell microarray can now be used for gene expression studies.
See our poster 'Bioreactor cultivation of CHO DP-12 cells under sodium butyrate treatment - comparative transcriptome analysis with CHO cDNA microarrays'by Klausing S, Krämer O and Noll T, presented at the ESACT conference 2011 for a first application. The poster can be found at the download area of our homepage.