Rice (oryza Sativa L.) R-pcd5, Gene Cloning And Characteristics Analysis And Fca Interacting Protein

In order to study the rice differentiating callus-specific expressed genes in Oryza sativa L., some important genes were screened by SSH (suppression subtractive hybridization).1. The Cloning and Characterzations of R-pcd5 Gene and Alternative splicing isoformAn EST fragment encoding partial rice PCD gene, had been obtained from SSH library. Using its translated amino acid sequence as query we searched GeneBank and found the gene homology to pcd5 programmed cell death 5 of Humo sapiens. Also several alternative splicing patterns of this gene were found in the EST database. So we designed two pairs of primers, obtained the complete cDNA and two alternative splicing cDNAs of the R-pcd5 gene by PCR and RT-PCR. Its complete cDNA including 414bp contains a 387 bp open reading frame and encodes a protein containing 128 amino acid protein.It expressed highly at transcriptional step in organs where PCD occurred actively, including old leaves, old roots, and the tissue where young panicle formed. It showed R-pcd5 played important roles on PCD. Quantitative PCR was performed to reveal that R-pcd5 was up-regulated in abiotic stress (low temperature and NaCl treatment). Neither R-pcd5 a nor R-pcd5 p includes a complete open reading frame and Both them encode truncated proteins, that expressed much lowier than R-pcd5 in mRNA level. 2 Detecting partner protein interacted with FCA.The rFCA homologous to Flowering-time controlling of Arabidopsis was cloned from rice by screening the SSH library, rFCA-1 is high homologous to FCA-gammaof Arabidopsis and contained conserved domains (two RNA-binding domains and a WW-domain). We tried to detect interaction protein of FCA via two-hybrid screening from rice, then the result show that the two positive-clone-sequences we have gotten are similarly to genomic fragments in Oryza sativa. The function of genes are further to be defined...