甘蔗蔗糖磷酸合成酶（SPS）基因克隆及其调控表达机制的研究

项目名称： 甘蔗蔗糖磷酸合成酶（SPS）基因克隆及其调控表达机制的研究

项目编号： No.30871575

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 陈由强

作者单位： 福建师范大学

项目金额： 34万元

中文摘要： 本项目以甘蔗为研究对象，克隆了表达蔗糖磷酸合成酶基因（SPSⅢ#65289;的甘蔗SPSⅢ#22522;因cDNA序列和DNA序列，研究了基因的结构并预测了功能；通过拟南芥异位表达，开展了甘蔗SPSⅢ#22522;因的过表达研究；研究克隆了甘蔗SPSⅢ#22522;因1.4Kb的5'侧翼序列，预测到大量与光响应有关的的顺式调节元件（如ACE 、ATCT-motif、AE-box等）和一些与组织发育相关的序列；研究开展了缺失表达分析，初步筛选出15个与其作用的特异转录因子；研究了SPSⅢ#22312;成熟期蔗茎中转录表达的规律，结合定量PCR和糖分检测，分析蔗茎中蔗糖积累与SPSⅢ#36716;录水平的相关性。研究发现并初步验证了甘蔗SPSⅢ#21069;体mRNA可变剪接体现象；研究还对甘蔗SPSⅢ#23478;族基因光暗磷酸化调控位点的机制开展研究。本项目的研究将为评估SPSⅢ#22522;因作为候选基因应用于甘蔗基因工程改良提供重要的理论参考。

中文关键词： 甘蔗；蔗糖磷酸合成酶；表达；转录

英文摘要： The DNA framents and cDNA framents coding entire SPSⅢwere isolated from sugarcane. The 5'-flanking sequence of SPSⅢwas cloned. These 5'-flanking sequence of SPSⅢwere fused to the coding Sequence of GUS gene in different length just to perform 5'-deletional analysis. It was observed that the gene GUS directed by the fragment downstream to position -725 expressed only in the base of receptacle and anther. It had been obtained 15 positive clones of different cDNA sequences by AbA screening .The five SPS gene families（SPSⅠ#12289;SPSⅡ#12289;SPSⅢ#12289;SPSⅣ#12289;SPSⅤ#65289; in the different tissues of sugarcane was determined by real-time PCR. During accumulate of sucrose, relative expression of SPS gene families had remained high level. Alternative splicing of SPS pre-mRNA was identified by RT-PCR in Saccharum.spp varieties. The hybridization signals furtherly showed that SPSⅢRNA had intron retention variants. But it was proved that the loss motif of TGCATG and TCTATC had no clear effect on the SPSⅢ9th exon normal splicing.

英文关键词： sugarcane;SPS;expression;transcription