Isolation and characterization of three families of auxin down-regulated cDNA clones
Isolation and characterization of three families of auxin down-regulated cDNA clones
复制标题
生长素下调 cDNA 克隆的三个家族的分离和表征
作者：
N. Datta;P. LaFayette;P. Kroner;R. T. Nagao;J. L. Key
分离并表征了代表三个生长素下调 (ADR) 基因家族的 5 个 cDNA 克隆（ADR6、ADR11-1、ADR11-2、ADR12-1 和 ADR12-2）。这些是通过用早期分离的部分 cDNA 克隆 p6、p11 和 p12 筛选 λZap cDNA 文库来分离的（Baulcombe 和 Key, J Biol Chem 255: 8907–8913, 1980）。当通过 SDS-PAGE 分析时，ADR6、ADR11-2 和 ADR12-2 克隆的杂交选择翻译分别产生 33 kDa、22.5 kDa 以及 6 和 7 kDa 的多肽。 ADR6 和 ADR12-2 在 IEF-SDS 2D 凝胶上分别产生 1 个和 2 个点。 ADR11-2 可能编码碱性蛋白，因为它只能通过非平衡 pH 梯度凝胶电泳 (NEPHGE) 才能解析。 ADR6、ADR11 和 ADR12 的基因组 Southern 印迹分析表明，每一个都代表一个小的多基因家族。与 ADR6、ADR11 和 ADR12 相对应的 RNA 水平响应应用的生长素而分别降低 100、15 和 10 倍（Baulcombe 和 Key，1980）。在存在和不存在生长素的情况下进行的径流转录表明，在存在生长素的情况下，与 ADR6、ADR11-2 和 ADR12-2 相对应的基因的转录速率降低，但相对于这些响应生长素的 mRNA 的细胞质水平的降低，这种降低幅度很小。生长素对体外转录和对应于这些 ADR cDNA 的稳态 RNA 水平的影响的比较分析表明，由于生长素引起的转录速率的降低不足以解释生长素诱导的稳态水平的降低。 Northern 分析显示了这些 ADR 基因的发育和器官/组织特异性反应。此外，对应于ADR6和ADR12-1的基因的表达似乎被光上调，而对应于ADR11的基因的表达似乎被光下调。
Five cDNA clones (ADR6, ADR11-1, ADR11-2, ADR12-1 and ADR12-2), representing three families of auxin down-regulated (ADR) genes were isolated and characterized. These were isolated by screening a λZap cDNA library with the partial cDNA clones p6, p11 and p12, isolated earlier (Baulcombe and Key, J Biol Chem 255: 8907–8913, 1980). Hybrid-select translation of ADR6, ADR11-2 and ADR12-2 clones produced polypeptides of 33 kDa 22.5 kDa and a 6 and 7 kDa respectively, when analyzed by SDS-PAGE. ADR6 and ADR12-2 gave one and two spots, respectively, on an IEF-SDS 2D gel. ADR11-2 probably encodes a basic protein as it was only resolved on non-equilibrium pH gradient gel electrophoresis (NEPHGE). Genomic Southern blot analysis of ADR6, ADR11 and ADR12 suggests that each represents a small multigene family. The RNA levels corresponding to ADR6, ADR11 and ADR12 decrease in response to applied auxin by 100-, 15- and 10-fold, respectively (Baulcombe and Key, 1980). Runoff transcription, done in the presence and absence of auxin, showed that the rate of transcription of the genes corresponding to ADR6, ADR11-2 and ADR12-2 was reduced in the presence of auxin, but the decrease was small relative to the decrease in the cytoplasmic levels of these mRNAs, in response to auxin. A comparative analysis of the influence of auxin on in vitro transcription and steady state RNA levels corresponding to these ADR cDNAs suggests that the decrease in rate of transcription due to auxin is not enough to account for the auxin-induced decrease in the steady state levels. Northern analysis showed developmental and organ/tissue-specific response of these ADR genes. Furthermore, the expression of the genes corresponding to ADR6 and ADR12-1 appears to be upregulated by light, whereas the gene corresponding to ADR11 appears to be down-regulated by light.
影响因子：
11.4
作者：
LUTCKE, HA;CHOW, KC;SCHEELE, GA
通讯作者：
SCHEELE, GA