Thornewell S J, Peery R B, Skatrud P L
Infectious Diseases Research, Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
Gene. 1997 Nov 12;201(1-2):21-9. doi: 10.1016/s0378-1119(97)00421-6.
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 approximately 152kDa. 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.
从新型隐球菌（Cn）临床分离株（菌株M1 - 106）中鉴定、克隆并表征了CneMDR1基因，该基因编码一种与几种真核生物多药耐药（MDR）蛋白相关的蛋白质。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CneMDR1。对相应cDNA的分析揭示了一个由16个内含子间断的开放阅读框。CneMDR1编码一种含有1408个氨基酸（aa）的蛋白质（CNEMDR1），预测分子量约为152kDa。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该Cn临床分离株中存在12条染色体带。通过在第四条染色体上杂交检测到CneMDR1。高严谨度杂交仅检测到一个MDR样基因。然而，在使用cDNA进行逆转录聚合酶链反应（RT-PCR）扩增过程中发现了第二个MDR样基因（CneMDR2）。
