Identification and localization of renal Na(+)-Ca2+ exchanger by polymerase chain reaction.
Identification and localization of renal Na(+)-Ca2+ exchanger by polymerase chain reaction.
复制标题
聚合酶链反应鉴定和定位肾Na()-Ca2交换体。
DOI：
10.1152/ajprenal.1992.263.4.f680
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发表时间：
1992
期刊：
影响因子：
--
通讯作者：
Lytton,J
中科院分区：
文献类型：
--
作者：
Yu,AS;Hebert,SC;Lee,SL;Brenner,BM;Lytton,J
采用基于同源性的聚合酶链反应（PCR）克隆策略确定了肾Na(+)-Ca2+交换器的分子特性。利用已发表的犬心脏Na(+)-Ca2+交换器cDNA序列中低退化区域的寡核苷酸引物，通过PCR扩增大鼠肾RNA，并对产物进行亚克隆和测序。鉴定出一个452 bp的克隆（NCX1），该克隆与犬心脏交换器有89%的核苷酸和98%的氨基酸序列相同，表明它们是同一基因的产物。Northern分析显示，NCX1与一个丰富的7 kb的主要转录本和一个大约14 kb的次要转录本杂交，这两个转录本主要定位于肾皮质。微解剖小管PCR分析显示，与其他皮质肾元段相比，NCX1在远曲小管中富集。这个位置与NCX1对应的Na(+)-Ca2+交换器在该位点的活性Ca2+重吸收中起主要作用是一致的。
The molecular identity of the renal Na(+)-Ca2+ exchanger was determined by a homology-based polymerase chain reaction (PCR) cloning strategy. Rat kidney RNA was amplified by PCR, using oligonucleotide primers based on regions of low degeneracy in the published canine cardiac Na(+)-Ca2+ exchanger cDNA sequence, and the products were subcloned and sequenced. A 452-bp clone (NCX1) was identified, which shares 89% nucleotide and 98% amino acid sequence identity with the canine cardiac exchanger, suggesting that they are products of the same gene. NCX1 was shown, by Northern analysis, to hybridize to an abundant major transcript of 7 kb and a minor one of approximately 14 kb both localized predominantly to kidney cortex. Microdissected tubule PCR analysis revealed that NCX1 was enriched in distal convoluted tubule compared with other cortical nephron segments. Such a location is consistent with a Na(+)-Ca2+ exchanger corresponding to NCX1 playing a major role in active Ca2+ reabsorption at this site.