Campos F, Carsolio C, Kuin H, Bisseling T, Rocha-Sosa M, Sánchez F
Departamento de Biología Molecular de Plantas, Universidad Nacional Autónoma de México, Cuernavaca.
Plant Physiol. 1995 Oct;109(2):363-70. doi: 10.1104/pp.109.2.363.
We previously reported that transcripts for a 30-kD nodulin (Npv30) are very abundant in the nodule. In this paper we describe the isolation and characterization of Npv30 cDNA and genomic clones. Npv30 has the following characteristic features: (a) a putative signal sequence at the deduced amino-terminal region, (b) a proline-rich stretch at the carboxy terminus, and (c) a characteristic domain of four cysteines that resemble metal-binding sites. In Phaseolus vulgaris L., Npv30 is encoded by a small gene family that shares discrete sequence homologies with another small gene family in soybean. An antibody against a beta-galactosidase-Npv30 fusion protein detected two proteins of 28 and 30 kD. Although Npv30 transcripts are very abundant, they encode proteins that are hardly detected in nodule fractions, suggesting that these proteins have a short half-life and/or the mRNAs are strongly regulated at the translational level. Npv30 transcripts were detected in the infected cells of the nodule by in situ hybridization experiments.
我们之前报道过,一种30-kD结节蛋白(Npv30)的转录本在根瘤中非常丰富。在本文中,我们描述了Npv30 cDNA和基因组克隆的分离与特性。Npv30具有以下特征:(a)在推导的氨基末端区域有一个假定的信号序列,(b)在羧基末端有一个富含脯氨酸的区域,以及(c)一个由四个半胱氨酸组成的特征结构域,类似于金属结合位点。在菜豆中,Npv30由一个小基因家族编码,该家族与大豆中的另一个小基因家族具有离散的序列同源性。一种针对β-半乳糖苷酶-Npv30融合蛋白的抗体检测到了两种分子量分别为28 kD和30 kD的蛋白质。尽管Npv30转录本非常丰富,但它们编码的蛋白质在根瘤组分中几乎检测不到,这表明这些蛋白质半衰期较短和/或mRNA在翻译水平受到强烈调控。通过原位杂交实验在根瘤的感染细胞中检测到了Npv30转录本。