Full-length CDNA Library Construction Of Rice Panicle At Early Milky Stage Under High Temperature Stress

In our previous study,54different expressing gene fragments and21different displaying functional proteins were identified by cDNA-AFLP genes different expressing technology and proteomics, respectively, however, the54different display genes and21different displaying functional proteins were not full length mRNA, In order to get full the length mRNA, Full-length cDNA library construction of rice panicle at early milky stage under high temperature stress. Thus, in this study, full length cDNAs were synthesized by the SMART PCR cDNA Synthesis Kit produced by Clontech company, with the mixed panicles of rice under high temperature stress and normal cultural conditions. After recovery and pure in sections, full length cDNA were ligated to T vector and transferred to DH5a, and7974positive clones were obtained. Among the clones, the size of target cDNA up to1.0kb were5074clones, account to63.6%. and the size of target cDNA down to1.0kb were2900clones, account to36.4%.Randomly selected34clones were sequenced, sequencing results in a Genbank EST database on rice. According to the result of random sequencing and blasting, the size of target cDNA up to1.0kb clones were the longest cDNA in the ESTs database, account75%. At the same time, comparison result display, genes OsP0003and OsP0012were highly homologous to the ESTs related to panicle fertile, and genes OsP0004, OsP0023, OsP0029and OsP0032were highly homologous to the ESTs related to drought stress. According to the quantifies and sequencing and blast results of clones, the library constructed in this study were included the major cDNA informations of rice gene expressing. The full length library provided a strict base for screening the full length cDNA of the different display genes involving in rice adapting to high temperature stress.