| Hypertension is one of the most prevalent cardiovascular diseases in the world, whose cases is increasing steadily in China. It is accepted that essential hypertension is the one that is associated with multiple genes. Research shows that almost 60% of the hypertensive patients are salt-sensitive. Na(+)-K(+)-Exchanging ATPase and Atrial Natriuretic Peptide (ANP) are the two candidate genes which were thought to be associated with salt sensitive hypertension.Na(+)-K(+)-Exchanging ATPase is a universal cell membrane protein in higher eukaryotic cells which mediates the anti-concentration gradient exchange of intracellular Na+ for extracellular K+. It plays essential metabolic roles such as maintaining sodium and potassium ion gradients across the plasma membrane. The targeted sequences were amplified by PCR to determine the genome structure of the hithertofore unsequenced portion of the alpha 1 subunit of the human Na(+)-K(+)-Exchanging ATPase gene which encodes 80-130 aa of its extracellular domain using two different templates, human genomic DNA and a human muscle cDNA library. The PCR products were analyzed by restriction endonuclease digestion and then cloned into a plasmid vector for chemiluminescence sequencing and further analysis. Two PCR products, 833bp (Fg) and 195bp (Fc) were isolated after PCR amplification with human genomic DNA and a muscle cDNA library, respectively. The size difference is accounted for by an insertion of a 638bp intron fragment in Fg at nt138 of Fc. A Blastn search of the nr Database in Genbank revealed no sequence homologous to the inserted fragment. A genomic fragment containing one complete human Na(+)-K(+)-Exchanging ATPase alpha 1 subunit gene intron was sequenced, and the splicing donor and receptor sites were identified. The accession number of this sequence is L76938. Analysis of this sequence shows the presence of several consensus sequences for transcription factor binding, such as CdxA, CP1.
|
PDF Full Text Request