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不同大小和丰度的器官特异性转录本源自玉米中同一个丙酮酸、正磷酸二激酶基因。

Organ-specific transcripts of different size and abundance derive from the same pyruvate, orthophosphate dikinase gene in maize.

作者信息

Glackin C A, Grula J W

机构信息

Phytogen, Pasadena, CA 91105.

出版信息

Proc Natl Acad Sci U S A. 1990 Apr;87(8):3004-8. doi: 10.1073/pnas.87.8.3004.

Abstract

Analyses of genomic DNA and clones indicate that the pyruvate, orthophosphate dikinase (PPDK; ATP: pyruvate, orthophosphate phosphotransferase, EC 2.7.9.1) gene family of maize (Zea mays L. subsp. mays, line B73) contains two members. Restriction site and DNA sequence comparisons between PPDK genomic and leaf cDNA clones have revealed which gene encodes the isozyme involved in C4 photosynthesis. The region flanking the 5' end of this gene contains two 30-base-pair (bp) repetitive elements that may be involved in its light-regulated expression. Sequence analysis of genomic and leaf cDNA clones has also shown that the entire 7.3-kDa PPDK chloroplast transit peptide is encoded in the 436-bp first exon. Northern blot experiments with probes specific for the first exon and the 3' end of the gene showed that the smaller PPDK transcripts in roots and etiolated leaves [3.0 kilobases (kb) vs. the 3.5-kb green leaf transcript] lack the sequence encoding the chloroplast transit peptide. In addition, results from cDNA library screens have confirmed that the root transcript is approximately 50-fold less abundant than the green leaf transcript. Finally, sequence comparisons among cDNA clones from green leaves and roots and genomic clones representing both members of the PPDK gene family demonstrate that the green leaf transcript encoding the C4 isozyme and the root transcript are derived from the same gene.

摘要

对基因组DNA和克隆的分析表明,玉米(Zea mays L. subsp. mays,B73品系)的丙酮酸,正磷酸二激酶(PPDK;ATP:丙酮酸,正磷酸磷酸转移酶,EC 2.7.9.1)基因家族包含两个成员。PPDK基因组克隆和叶片cDNA克隆之间的限制性酶切位点和DNA序列比较揭示了哪个基因编码参与C4光合作用的同工酶。该基因5'端侧翼区域包含两个30碱基对(bp)的重复元件,可能参与其光调控表达。基因组和叶片cDNA克隆的序列分析还表明,整个7.3-kDa的PPDK叶绿体转运肽由436-bp的第一个外显子编码。用针对该基因第一个外显子和3'端的探针进行的Northern印迹实验表明,根和黄化叶片中较小的PPDK转录本[3.0千碱基(kb),而绿色叶片转录本为3.5-kb]缺乏编码叶绿体转运肽的序列。此外,cDNA文库筛选结果证实,根转录本的丰度比绿色叶片转录本低约50倍。最后,来自绿色叶片和根的cDNA克隆以及代表PPDK基因家族两个成员的基因组克隆之间的序列比较表明,编码C4同工酶的绿色叶片转录本和根转录本来自同一个基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/448c/53822/ed88849d689b/pnas01033-0148-a.jpg

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