Isolation and characterization of a sodium-dependent phosphate transporter gene in Dunaliella viridis
Isolation and characterization of a sodium-dependent phosphate transporter gene in Dunaliella viridis
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DOI：
10.1016/j.bbrc.2005.11.144
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发表时间：
2006-02-03
影响因子：
3.1
通讯作者：
Xu, ZK
中科院分区：
文献类型：
--
作者：
Li, QY;Gao, XS;Xu, ZK
利用从绿色杜氏藻消减文库中提取的探针，从文库中克隆到一个钠依赖的磷酸转运蛋白基因DvSPT1。测序结果显示，该基因全长2649bp，编码672个氨基酸的开放阅读框。DvSPT1的氨基酸序列与Tetraselmis Chui的TcPHO有31.2%的同源性。疏水性和二级结构预测揭示了11个保守的跨膜结构域，与酿酒酵母的PHO89和粗糙脉孢霉的PHO4相似。Northern印迹分析表明，DvSPTI在高渗胁迫或磷耗竭条件下被诱导表达。酵母Na+输出泵突变体G19的功能鉴定表明，DvSPT1编码一个Na+转运蛋白。从绿色木霉基因组文库中克隆了GDvSPT1基因序列(7922bp)。Southern杂交分析表明，绿色假丝酵母中至少存在两个同源基因。(C)2005 Elsevier Inc.保留所有权利。
A sodium-dependent phosphate transporter gene, DvSPT1, was isolated from a cDNA library using a probe derived from a subtracted cDNA library of Dunaliella viridis. Sequencing analyses revealed a cDNA-sequence of 2649 bp long and encoded an open-reading frame consisting of 672 amino acids. The deduced amino acid sequence of DvSPT1 exhibited 31.2% identity to that of TcPHO from Tetraselmis chui. Hydrophobicity and secondary structure prediction revealed 11 conserved transmembrane domains similar to those found in PHO89 from Saccharomyces cerevisiae and PHO4 from Neurospora crassa. Northern blot analysis indicated that the DvSPTI expression was induced upon NaCl hyperosmotic stress or phosphate depletion. Functional characterization in yeast Na+ export pump mutant G19 suggested that DvSPT1 encoded a Na+ transporter protein. The gene sequence of GDvSPT1 (7922 bp) was isolated from a genomic library of D. viridis. Southern blot analysis indicated that there exist at least two homologous genes in D. viridis. (c) 2005 Elsevier Inc. All rights reserved.