Tingey S V, Walker E L, Coruzzi G M
EMBO J. 1987 Jan;6(1):1-9. doi: 10.1002/j.1460-2075.1987.tb04710.x.
We have characterized the distinct polypeptides, primary translation products and mRNAs encoding glutamine synthetase (GS) in the various organs of pea. Western blot analysis of soluble protein has identified five distinct GS polypeptides which are expressed at different relative levels in leaves, roots and nodules of pea. Of the two GS polypeptides in leaves (44 and 38 kd), the 44-kd GS polypeptide is predominant and is localized to the chloroplast stroma. In roots, the predominant GS polypeptide is 38 kd. Upon Rhizobium infection of roots, three 37-kd GS polypeptides increase in abundance in the nodules relative to uninfected roots. cDNA clones encoding three different GS mRNAs have been characterized. Hybrid-select translation has identified three different GS primary translation products (49, 38 and 37 kd). Two cDNA clones (pGS134 and pGS341) are homologous to GS mRNAs most abundant in nodules which encode the 38- and 37-kd GS primary translation products. A third cDNA (pGS197) corresponds to a larger GS mRNA species specific to leaf poly(A) RNA, which encodes a 49-kd putative precursor to the mature chloroplast GS polypeptide. cDNA sequence analysis and Southern blot analysis of pea nuclear DNA identifies at least three genes encoding GS in pea which are related but distinct in structure and in vivo pattern of expression.
我们已对豌豆各器官中编码谷氨酰胺合成酶(GS)的不同多肽、初级翻译产物和mRNA进行了表征。对可溶性蛋白的蛋白质免疫印迹分析确定了五种不同的GS多肽,它们在豌豆的叶片、根和根瘤中以不同的相对水平表达。在叶片中的两种GS多肽(44 kd和38 kd)中,44 kd的GS多肽占主导地位,定位于叶绿体基质。在根中,占主导地位的GS多肽是38 kd。根被根瘤菌感染后,相对于未感染的根,根瘤中三种37 kd的GS多肽丰度增加。已对编码三种不同GS mRNA的cDNA克隆进行了表征。杂交选择翻译确定了三种不同的GS初级翻译产物(49 kd、38 kd和37 kd)。两个cDNA克隆(pGS134和pGS341)与根瘤中最丰富的GS mRNA同源,它们编码38 kd和37 kd的GS初级翻译产物。第三个cDNA(pGS197)对应于一种较大的GS mRNA,它是叶片多聚腺苷酸化RNA特有的,编码成熟叶绿体GS多肽的49 kd推定前体。豌豆核DNA的cDNA序列分析和Southern印迹分析确定豌豆中至少有三个编码GS的基因,它们在结构和体内表达模式上相关但不同。