首页> 美国卫生研究院文献>Nucleic Acids Research >Cloning of cDNA for pyruvate Pi dikinase from maize leaves.
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Cloning of cDNA for pyruvate Pi dikinase from maize leaves.

机译:玉米叶片丙酮酸Pi二激酶的cDNA的克隆。

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摘要

To obtain molecular probes for studies of gene regulation in photosynthetic tissues of maize, we have cloned DNA complementary to poly(A)+RNA extracted from green leaves by insertion into plasmid pBR322 and transformation of E. coli, strain RR1. Colonies were screened by sequential hybridization with 32P-labeled single stranded cDNA synthesized from pooled aliquots of poly(A)+RNA fractionated by sucrose density centrifugation. Among the clones bearing cDNA homologous to high molecular weight poly(A)+RNA, we identified one with an insert of 440 base pairs homologous to mRNA for pyruvate, Pi dikinase, a C-4 carbon cycle protein localized in mesophyll cells of the leaf. Our work indicates that the dikinase subunits are synthesized in the cytoplasm as precursors approximately 13,000 daltons larger than the mature peptide subunits. Leaves of seedlings illuminated during growth have higher levels of pyruvate, Pi dikinase mRNA than leaves of dark-grown plants.
机译:为了获得用于研究玉米光合组织中基因调控的分子探针,我们通过插入质粒pBR322并转化大肠杆菌RR1菌株,克隆了与从绿叶中提取的poly(A)+ RNA互补的DNA。通过与由蔗糖密度离心分离的聚(A)+ RNA的合并等分试样合成的32P标记单链cDNA进行顺序杂交,筛选菌落。在携带与高分子量poly(A)+ RNA同源的cDNA的克隆中,我们鉴定出一个插入有440个碱基对的插入物,与丙酮酸,Pi dikinase,一种位于叶肉叶肉细胞中的C-4碳循环蛋白的mRNA同源。 。我们的工作表明,二激酶亚基在细胞质中合成为前体,比成熟的肽亚基大约13,000道尔顿。生长期间照亮的幼苗叶片的丙酮酸,Pi二激酶mRNA水平高于深色植物的叶片。

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