Summary: | 碩士 === 國立中興大學 === 生物科技學研究所 === 98 === Eukaryotic gene expression is a complex process includes transcriptional, post-transcriptional (e.g. pre-mRNA processing), translational, and post-translational regulations (e.g. protein modifications). During pre-mRNA processing event, polyadenylation is mediated by adding polyadenosine into newly formed 3’ end of mRNA after nascent mRNA cleavage, but its molecular mechanism is still unclear in plant. However, recent studies indicated that the nascent mRNA cleavage is mediated by a novel cleavage stimulation factor (CstF) complex that does not directly cleavage RNA but assists other proteins to process RNA through recognizing specific cis-element in nascent mRNA itself. Our long-termed interest is to identify and functionally investigate the genes/proteins involved in pollen development and pollination. By taking advantage of tremendous resources generated from the studies of Arabidopsis, we used bioinformatics and data-mining approach at the genome level to search the T-DNA insertion mutants with defective male gametophyte development. One of mutants that could not produce homozygotic progeny is caused by T-DNA insertional disruption of Arabidopsis gene Non Homozygotic Progeny 25 (NHP25), which was confirmed by flanking sequencing. Interestingly, among all the characterized Arabidopsis T-DNA line of NHP25, two of them (nhp25-1 and nhp25-5) might be leakage and showed pleiotropic phenotypes in sterile homozygote mutants including late flowering, draft growth, jigsaw rosette leave, abnormal flower morphology, and seed abortion. Complementary assay shows fully complement in two different T-DNA insertion alleles. The microarray experiment surveyed the genes which are affected in nhp25 mutant and found two flowing time regulated genes, SPL3 and SPL4 are significantly downregulated in nhp25 mutant, suggesting the NHP25 might regulate flowering time through controlling SPL3 and SPL4 expression.
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