Project description:Identification of new and unpredicted full length Arabidopsis genes. Examination of cRNA prepared from Arabidopsis thaliana ecotype Columbia light grown 7-day old seedlings using whole genome tiling arrays. Keywords: other
Project description:Identification of new and unpredicted full length Arabidopsis genes. Examination of cRNA prepared from Arabidopsis thaliana ecotype Columbia light grown 7-day old seedlings, light grown 7-day old seedlings treated at 4 degrees C for 24hrs, and etiolated 7-day old seedlings using pilot genome tiling arrays. Total RNA was isolated from seedlings with the TRIzol reagent procedure of Invitrogen, then poly(A)+ RNA was purified with Qiagen oligotex. One microgram poly(A)+ RNA was converted into ds-cDNA using T7-oligo(dT) primers. Biotin-labeled cRNA was generated by in vitro transcription reactions using ENZO labeling kit, fragmented and then 20 micrograms of fragmented cRNA were hybridized to the arrays according to Affymetrix instructions. Keywords: other
Project description:Examination of cRNA from Arabidopsis flowers, roots and suspension cell culture using custom high-density Affymetrix genome tiling arrays in order to identify new and unpredicted genes. Keywords: other
Project description:In this study, we used Arabidopsis root extracts, spiked with amide nitrogen labeled (15N1) Glutamine and a purified recombinant protein, both full length and glutaminase domain only versions, to determine the amido group acceptor, if any, in the glutamine amidotransferase reaction.
Project description:The length of Arabidopsis microRNA genes and their precursors (pri-miRNAs) can very between several hundreds bp to several thousands bp. They may contain introns, precursors may undergo alternative splicing, and often there are more than one polyadenylation site. An intriguing question arise: why to transcribe thousands of base pairs to produce a small RNA molecule of about 21 nt in length? We decided to sequence small RNAs from 35-day-old rosette leaves and to map the obtained reads of 18-26 nt in length to Arabidopsis pri-miRNAs. Our analyses revealed the presence of sRNAs that can derive only from miRNA precursors. An example is the identification of a substantial number of miR319b.2 reads. This sRNA is a product of pri-miR319b processing. Our further experimental analyses show that it carries multiple signatures of a functional microRNA
