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The cloning genetic mapping and expression of the constitutive sucrose synthase locus of maize

机译:玉米组成型蔗糖合酶基因座的克隆遗传定位及表达

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摘要

Two differentially expressed genes encode isoenzymes of sucrose synthase in Zea mays. A clone of the shrunken 1 (Sh1) locus, the structural gene for the major endosperm form of sucrose synthase, was used to isolate a genomic clone of constitutive sucrose synthase (Css), the structural gene for the isoenzyme expressed in embryo and other tissues. The Css clone was positively identified by RNA blot analysis of RNA from wild type and a sh1 deletion stock and by analysis of the in vitro translation product of hybrid-selected mRNA. Southern blot analysis of DNA from monosomic plants derived from an r-x1 stock, coupled with restriction fragment length polymorphism mapping, placed the Css gene 32 map units from Sh1 on chromosome 9. In seedling tissues, Css mRNA is present at higher levels than Sh1 mRNA. Expression of both Sh1 and Css in root tissue is enhanced by anaerobic conditions, although Css is induced to a lesser extent than is Sh1. Thus, Css appears to be expressed constitutively, whereas Sh1 is expressed at high levels only in response to specific developmental and environmental stimuli.
机译:两个差异表达的基因编码玉米中蔗糖合酶的同工酶。收缩的1(Sh1)基因座(一种主要的胚乳形式的蔗糖合酶的结构基因)的克隆用于分离组成型蔗糖合酶(Css)的基因组克隆,该结构基因是在胚芽和其他组织中表达的同工酶的结构基因。通过对野生型和sh1缺失原种的RNA进行RNA印迹分析,以及对杂种选择的mRNA的体外翻译产物进行分析,可以正确鉴定Css克隆。对来自r-x1种群的单体植物的DNA进行Southern印迹分析,结合限制性片段长度多态性图谱,将Shs的Css基因32个图谱单元置于9号染色体上。在幼苗组织中,Css mRNA的表达水平高于Sh1。 mRNA。尽管Css的诱导程度小于Sh1,但厌氧条件可增强根组织中Sh1和Css的表达。因此,Css似乎是组成性表达,而Sh1仅在响应特定的发育和环境刺激时才以高水平表达。

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