水稻腊质基因分子特性的研究

水稻腊质基因(Wx)负责胚乳与花粉粒中直链淀粉的合成。我们通过对限制图和DNA顺序有重叠的两个基因组克隆的分析,测定出了水稻Wx基因全长为5499bp的DNA顺序。比较水稻、玉米(Klsgen等)和大麦(Rohde等)中Wx基因的DNA顺序,弄清水稻Wx基因中存在13个内含子和14个外显子,通过微机对外显子顺序的翻译,得出了水稻Wx蛋白包括转运肽在内的609个氨基酸的排列顺序,并计算出它的分子量约为72kD。经比较,水稻Wx成熟蛋白的氨基酸顺序与玉米、大麦的没有明显地差异。但是,水稻Wx基因5′-上游区、3′-下游区和内含子区域的DNA顺序,以及Wx前体蛋白的转运肽区域的氨基酸顺序与玉米和大麦Wx基因的相应区域相比,它们之间的同源性却很低。     

Identification of Two Transposon-like Elements in Rice Wx Gene

Rice Wx genes have been cloned and sequenced from Oryza saliva subsp. japonica (Wx-R-jp) and subsp. indica (Wx-R-id) and wild rice Oryza saliva L. f. spontanea (Wx-R-sp). Comparison of the nucleotide sequences of these three Wx genes indicates that Wx-R-jp and Wx-R-id genes contain two types of transposon-like elements, designated by RTL-1 and RTL-2. in the region of their two introns and 5' upstream, RTL-1 shares sequence homology with valine tRNA molecules, and contains an internal promoter of RNA polymerase Ⅲ. RTL-2 forms a stem-loop structure. Both RTL-1 and RTL-2 are flanked by direct repetitive sequences. Compared with the elements that have been known transposable, RTL-1 resembles the short interspersed-repeated DNA elements (SINEs) and RTL-2 is similar to the fold-back (FB) element of trahsposon of Drosophila.     

MOLECULAR CHARACTERIZATION OF RICE Wx GENE

The complete nucleotide (nt) sequence of the rice waxy(Wx) gene, which is responsible for the synthesis of amylose in endosperm and pollen, has been determined by a combination of restriction mapping and nt sequence analysis of two overlapping genomic DNA clones. The entire gene is about 5.5 kb in length. The alignment of the nt sequence of the Wx gene from rice with those of maize (Klsgen, R. B. et al.) and barley (Rohde, W. et al.) revealed the presence of thirteen introns and fourteen exons. The full-length of Wx protein in cluding transit peptide is 609 amino acid (aa) residues. The calculated molecular weight of rice Wx preprotein is about 72 kD. There is no significant difference between the similarity scores of the aa sequence deduced from the rice Wx gene compared with those of maize and barley. However, the nt sequences of the 5'-end upstream, 3'-end downstream and introns of the rice Wx gene, as well as the aa sequence of the transit peptide region of the Wx preprotein have low similarity scor     

水稻蜡质基因中两种类似转座因子的顺序

本实验从粳稻、籼稻与野生稻中分别克隆出了它们的蜡质基因(Wx-R-jp,Wx-R-id与Wx-R-sp),并且测定了它们的全顺序。将这3个蜡质基因的核苷酸顺序进行比较,结果表明在粳稻与籼稻蜡质基因的两个内含子与5′上游区中含有两种类似转座因子的结构,称之为RTL-1和RTL-2。RTL-1与缬氨酸tRNA基因及RNA多聚酶Ⅲ内部启动子顺序有一定程度的同源性。RTL-2能形成“茎环”结构。邻接RTL-1或RTL-2的两端都有短的顺向重复顺序。从分子结构上推测RTL-1类似于哺乳动物中的转座因子SINE (Short Interspersed Repeated DNA Elements),而RTL-2则与果蝇中的转座因子FB(foldback)有许多相似之处。