Shen B, Carneiro N, Torres-Jerez I, Stevenson B, McCreery T, Helentjaris T, Baysdorfer C, Almira E, Ferl R J, Habben J E
Dept. of Plant Sciences, University of Arizona, Tucson 85721.
Plant Mol Biol. 1994 Nov;26(4):1085-101. doi: 10.1007/BF00040691.
As one component of a maize genome project, we report the analysis of a number of randomly selected cDNAs, by a combination of measuring mRNA expression, 'single-pass' sequencing (SPS), and genome mapping. Etiolated seedling (490) and membrane-free polysomal endosperm cDNA clones (576) were evaluated for their transcription levels by hybridizing with a probe prepared from total mRNA and categorized as corresponding to abundantly or rarely expressed mRNAs and as either constitutive or tissue-specific. A total 313 clones from the two libraries were submitted to 'single-pass' sequencing from the presumed 5' end of the mRNA and the nucleotide sequence compared with the GenBank database. About 61% of the clones showed no significant similarities within GenBank, 14% of the clones exhibited a high degree of similarity, while the remaining 25% exhibited a lesser degree of similarity. The chromosomal location of more than 300 clones was determined by RFLP mapping using standard populations. The results demonstrate that a combination of analyses provides synergistic information in eventually deducing the actual function of these types of clones.
作为玉米基因组计划的一个组成部分,我们报告了通过测量mRNA表达、“单通道”测序(SPS)和基因组作图相结合的方法,对一些随机选择的cDNA进行的分析。通过与从总mRNA制备的探针杂交,评估黄化幼苗(490个)和无膜多聚核糖体胚乳cDNA克隆(576个)的转录水平,并将其分类为对应于高丰度或低丰度表达的mRNA,以及组成型或组织特异性的mRNA。来自两个文库的总共313个克隆从mRNA的假定5'端进行“单通道”测序,并将核苷酸序列与GenBank数据库进行比较。约61%的克隆在GenBank中未显示出显著相似性,14%的克隆表现出高度相似性,其余25%表现出较低程度的相似性。使用标准群体通过RFLP作图确定了300多个克隆的染色体位置。结果表明,这些分析方法相结合,在最终推断这类克隆的实际功能时能提供协同信息。