Belanger F C, Kriz A L
Department of Agronomy, University of Illinois, Urbana, Illinois, 61801.
Plant Physiol. 1989 Oct;91(2):636-43. doi: 10.1104/pp.91.2.636.
One of the most abundant proteins in maize (Zea mays L.) embryos is the molecular weight 63,000 globulin encoded by the Glb1 gene. To obtain DNA clones corresponding to Glb1, a cDNA library corresponding to RNA from developing maize embryos was constructed in a lambda expression vector and screened with antibodies specific for Glb1-encoded proteins. Here we report the complete nucleotide sequence, as determined from two overlapping clones, of pcGlb 1S, a 2009 base pair clone containing the entire translated region of Glb1. The deduced amino acid sequence of pcGlb 1S shows similarities to 7S-type seed storage proteins of wheat and legumes. Southern blot analysis of maize DNA confirms previous genetic studies which had indicated the presence of a single copy of Glb1 per haploid genome. Northern blot analysis indicates that Glb1 transcripts are present throughout most of embryo development and that expression of this gene is limited to seed tissues. Embryos homozygous for a Glb1 null allele, in which Glb1-encoded proteins are not detectable, contain low levels of Glb1 transcripts which are a different size from those encoded by functional alleles. This suggests that the defect in the null allele is at the level of gene transcription or RNA processing.
玉米(Zea mays L.)胚中最丰富的蛋白质之一是由Glb1基因编码的分子量为63,000的球蛋白。为了获得与Glb1对应的DNA克隆,在λ表达载体中构建了一个与发育中的玉米胚RNA对应的cDNA文库,并用对Glb1编码蛋白具有特异性的抗体进行筛选。在此,我们报告了pcGlb 1S的完整核苷酸序列,该序列由两个重叠克隆确定,pcGlb 1S是一个2009碱基对的克隆,包含Glb1的整个翻译区域。pcGlb 1S推导的氨基酸序列与小麦和豆类的7S型种子贮藏蛋白具有相似性。玉米DNA的Southern印迹分析证实了先前的遗传学研究,该研究表明每个单倍体基因组中存在一个Glb1拷贝。Northern印迹分析表明,Glb1转录本在胚发育的大部分时间都存在,并且该基因的表达仅限于种子组织。对于Glb1无效等位基因纯合的胚,其中检测不到Glb1编码的蛋白,含有低水平的Glb1转录本,其大小与功能等位基因编码的转录本不同。这表明无效等位基因中的缺陷在于基因转录或RNA加工水平。