Cloning and characterization of CneMDR1:: a Cryptococcus neoformans gene encoding a protein related to multidrug resistance proteins
Cloning and characterization of CneMDR1:: a Cryptococcus neoformans gene encoding a protein related to multidrug resistance proteins
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DOI：
10.1016/s0378-1119(97)00421-6
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发表时间：
1997-11-12
期刊：
影响因子：
3.5
通讯作者：
Skatrud, PL
中科院分区：
文献类型：
--
作者：
Thornewell, SJ;Peery, RB;Skatrud, PL
CneMDR 1是一个编码与几种真核多药耐药（MDR）蛋白相关的蛋白的基因，从新生隐球菌（Cn）（菌株M1-106）的临床分离株中鉴定、克隆和表征。最初使用聚合酶链反应（PCR）扩增包含其他MDR样蛋白保守基序的DNA区域来鉴定和克隆CneMDR 1。相应的cDNA的分析揭示了一个由16个内含子打断的开放阅读框架。CneMDR 1编码一个蛋白质（CNEMDR 1），含有1408个氨基酸，分子量约为152 kDa。蛋白质结构预测表明存在两个假定的6-跨膜（TM）结构域以及两个ATP结合结构域，这是ATP结合盒（ABC）蛋白的典型结构特征。这个超家族的成员，包括MDR蛋白，经常参与多种底物跨细胞膜的主动转运。脉冲场凝胶电泳显示存在12条染色体带在此临床分离的Cn。CneMDR 1通过IV号染色体上的杂交检测。高严谨性杂交仅检测到一个MDR样基因。然而，第二个MDR样基因（CneMDR 2）被发现在使用cDNA的逆转录酶-PCR（RT-PCR）扩增。(C)1997年Elsevier Science B.V.
CneMDR1, a gene encoding a protein related to several eukaryotic multidrug resistance (MDR) proteins, was identified, cloned, and characterized from a clinical isolate of Cryptococcus neoformans (Cn) (strain M1-106). Polymerase chain reaction (PCR) amplification of a DNA region encompassing conserved motifs of other MDR-like proteins was initially used to identify and clone CneMDR1. Analysis of the corresponding cDNA revealed an open reading frame punctuated by 16 introns. CneMDR1 encoded a protein (CNEMDR1) containing 1408 amino acids (aa) with a predicted mass of similar to 152 kDa. Protein structure predictions suggested the presence of two putative 6-transmembrane (TM) domains as well as two ATP-binding domains, structural characteristics typical of ATP-binding cassette (ABC) proteins. Members of this superfamily, which include MDR proteins, are frequently involved in active transport of a variety of substrates across the cell membrane. Pulsed-field gel electrophoresis revealed the presence of 12 chromosomal bands in this clinical isolate of Cn. CneMDR1 was detected by hybridization on chromosome IV. High-stringency hybridization detected only one MDR-like gene. However, a second MDR-like gene (CneMDR2) was discovered during reverse transcriptase-PCR (RT-PCR) amplification using cDNA. (C) 1997 Elsevier Science B.V.