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[Acquirement and identification of a novel intron 1 of human Na(+)-K(+)-exchanging ATPase alpha 1 subunit gene].

OBJECTIVE
The unknown part genome structure of the alpha 1 subunit of human Na(+)-K(+)-Exchanging ATPase gene which covers the 80-130 aa in its extracellular domain was studied.


METHODS
By means of PCR, the targeted sequences was amplified by using two different templates: human Genome DNA and the muscle cDNA library; the PCR products were analyzed by restriction endonuclease digestion, then cloned into plasmid vector to perform the chemiluminescence Sequencing. The homologous search, splicing site detection and the analysis of the sequence were applied.


RESULTS
Two different lengths of products, 833 bp(Fg) and 195 bp(Fc) were acquired after PCR amplification with the human genome DNA and muscle cDNA library respectively. There was an insertion of a 638 bp fragment in Fg at its 138-775 bp. And there was not obviously known sequence homologous to the insertion sequence.


CONCLUSIONS
A novel sequence containing the complete human Na(+)-K(+)-Exchanging ATPase alpha 1 subunit gene intron 1 was found, the exact splicing donor and receptor site within it were identified. The Accession No of this sequence is L76938. Analysis of this sequence showed that the intron 1 may function in the regulation of the gene expression and also had a potential coding sequence.