Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340.
Plant Physiol. 1986 Jun;81(2):642-5. doi: 10.1104/pp.81.2.642.
Western blots of soluble protein from wheat, rice, and corn showed that ADPglucose pyrophosphorylase subunits have a size of 50 kilodaltons from endosperm tissue and 43 and 46 kilodaltons from leaf. Antisera to ADPglucose pyrophosphorylase precipitated in vitro translation products of 73 and 76 kilodaltons when leaf poly(A)(+) RNA was used, whereas endosperm mRNA directed the synthesis of 50 and 56 kilodalton polypeptides. To further study the nature of these mRNA species, an ADPglucose pyrophosphorylase cDNA clone from rice endosperm polyadenylated RNA was obtained and used as a hybridization probe. Northern blots showed that ADPglucose pyrophosphorylase mRNA was slightly larger in leaf (2100 bases) than in endosperm tissue (1900 bases). These studies indicated that in cereals there are at least two tissue specific forms of ADPglucose pyrophosphorylase that are encoded by distinct mRNA transcripts. Analysis of genomic DNA by Southern blotting suggested that ADPglucose pyrophosphorylase is encoded by a small gene family.
来自小麦、水稻和玉米胚乳组织可溶性蛋白的 Western blot 分析表明,ADP-葡萄糖焦磷酸化酶亚基的大小为 50kDa;而来自叶片的大小为 43kDa 和 46kDa。当使用叶片 poly(A)(+) RNA 时,ADP-葡萄糖焦磷酸化酶抗血清沉淀体外翻译产物为 73kDa 和 76kDa,而胚乳 mRNA 则指导合成 50kDa 和 56kDa 多肽。为了进一步研究这些 mRNA 种类的性质,从水稻胚乳多聚 A RNA 中获得了 ADP-葡萄糖焦磷酸化酶 cDNA 克隆,并将其用作杂交探针。Northern blot 分析表明,ADP-葡萄糖焦磷酸化酶 mRNA 在叶片中略大于胚乳组织(2100 个碱基)(1900 个碱基)。这些研究表明,在谷物中,至少有两种组织特异性形式的 ADP-葡萄糖焦磷酸化酶,它们由不同的 mRNA 转录本编码。Southern blot 分析基因组 DNA 表明,ADP-葡萄糖焦磷酸化酶由一个小基因家族编码。