Identification of Copper-Induced Genes in the Marine Alga Ulva compressa (Chlorophyta)
Identification of Copper-Induced Genes in the Marine Alga Ulva compressa (Chlorophyta)
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DOI：
10.1007/s10126-010-9325-8
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发表时间：
2011-06-01
影响因子：
3
通讯作者：
Moenne, Alejandra
中科院分区：
文献类型：
--
作者：
Contreras-Porcia, Loretto;Dennett, Geraldine;Moenne, Alejandra
为了鉴定涉及铜耐受性的基因/蛋白质，用10 μ M铜培养海洋石莼3天。抗氧化酶抗坏血酸过氧化物酶（AP），过氧化物酶（PRX），硫氧还蛋白（TRX），谷胱甘肽-S-转移酶（GST）的活性和脂质过氧化物的水平进行了测定，在培养的蚯蚓与铜添加。抗氧化酶活性和过氧化脂质水平增加响应铜过量，表明在氧化应激。用U.用10 μ M铜培养3天。共分离出3 × 104个克隆，对480个克隆进行测序，得到235个非冗余EST，其中104个编码具有已知功能的蛋白质。其中，我们鉴定了参与（1）抗氧化代谢的蛋白质，如AP、PRX、TRX、GST和金属硫蛋白（MET），（2）信号转导，如钙调蛋白（CAM），（3）钙依赖性蛋白激酶（CDPK）和核苷二磷酸激酶（NDK），（4）基因表达，（5）蛋白质合成和降解，以及（6）叶绿体和线粒体电子传递链。鉴定的蛋白质中有一半可能定位于细胞器中。用定量RT-PCR方法测定了10 μ M铜处理0 ~ 7 d的黄瓜中AP、PRX、TRX、GST、MET、CAM、CDPK和NDK等18个基因的相对表达量。转录水平增加，响应铜胁迫，其中大部分在第3和第5天达到最大值。因此，筛选出的基因是受铜胁迫诱导的，可能与铜胁迫的适应和耐性有关。
In order to identify genes/proteins involved in copper tolerance, the marine alga Ulva compressa was cultivated with 10 mu M copper for 3 days. The activities of antioxidant enzymes ascorbate peroxidase (AP), peroxiredoxin (PRX), thioredoxin (TRX), and glutathione-S-transferase (GST) and the level of lipoperoxides were determined in the alga cultivated with and without copper addition. Antioxidant enzyme activities and lipoperoxides level increased in response to copper excess, indicating that the alga was under oxidative stress. A cDNA library was prepared using U. compressa cultivated with 10 mu M copper for 3 days. A total of 3 x 10(4) clones were isolated and 480 clones were sequenced, resulting in 235 non-redundant ESTs, of which 104 encode proteins with known functions. Among them, we identified proteins involved in (1) antioxidant metabolism such as AP, PRX, TRX, GST, and metalothionein (MET), (2) signal transduction, such as calmodulin (CAM), (3) calcium-dependent protein kinase (CDPK) and nucleoside diphosphate kinase (NDK), (4) gene expression, (5) protein synthesis and degradation, and (6) chloroplast and mitochondria electron transport chains. Half of the identified proteins are potentially localized in organelles. The relative level of 18 genes, including those coding for AP, PRX, TRX, GST, MET, CAM, CDPK, and NDK were determined by quantitative RT-PCR in the alga cultivated with 10 mu M copper for 0 to 7 days. Transcript levels increased in response to copper stress and most of them reached a maximum at days 3 and 5. Thus, the selected genes are induced by copper stress and they are probably involved in copper acclimation and tolerance.