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Human placental Na+,K+-ATPase alpha subunit: cDNA cloning, tissue expression, DNA polymorphism, and chromosomal localization.

机译:人胎盘Na +,K + -ATPaseα亚基:cDNA克隆,组织表达,DNA多态性和染色体定位。

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摘要

A 2.2-kilobase clone comprising a major portion of the coding sequence of the Na+,K+-ATPase alpha subunit was cloned from human placenta and its sequence was identical to that encoding the alpha subunit of human kidney and HeLa cells. Transfer blot analysis of the mRNA products of the Na+,K+-ATPase gene from various human tissues and cell lines revealed only one band (approximately 4.7 kilobases) under low and high stringency washing conditions. The levels of expression in the tissues were intestine greater than placenta greater than liver greater than pancreas, and in the cell lines the levels were human erythroleukemia greater than butyrate-induced colon greater than colon greater than brain greater than HeLa cells. mRNA was undetectable in reticulocytes, consistent with our failure to detect positive clones in a size-selected (greater than 2 kilobases) lambda gt11 reticulocyte cDNA library. DNA analysis revealed a polymorphic EcoRI band and chromosome localization by flow sorting and in situ hybridization showed that the alpha subunit is on the short arm (band p11-p13) of chromosome 1.
机译:从人胎盘克隆出包含Na +,K + -ATPaseα亚基编码序列主要部分的2.2碱基碱基克隆,其序列与编码人肾脏和HeLa细胞的α亚基的序列相同。来自各种人体组织和细胞系的Na +,K + -ATPase基因的mRNA产物的转移印迹分析表明,在低和高严格洗涤条件下,只有一个谱带(约4.7千碱基)。组织中的表达水平是小肠大于胎盘大于肝大于胰腺,并且在细胞系中的表达水平是人红白血病大于丁酸盐诱导的结肠大于结肠大于脑大于HeLa细胞。在网织红细胞中无法检测到mRNA,这与我们未能在大小选择的(大于2 KB)λgt11网织红细胞cDNA文库中检测到阳性克隆失败有关。 DNA分析显示多态性EcoRI条带,通过流分选和原位杂交显示染色体定位表明,α亚基位于1号染色体的短臂(p11-p13条带)上。

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