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Cloning of the rat heart Na + ‐Ca 2+ exchanger and its functional expression in HeLa cells
Author(s) -
Low Walter,
Kasir Judith,
Rahamimoff Hannah
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)81737-k
Subject(s) - cloning (programming) , hela , microbiology and biotechnology , chemistry , biology , cell culture , genetics , computer science , programming language
A functional rat heart Na + ‐Ca 2+ exchanger gene has been obtained by splicing and ligating two partially overlapping clones isolated from a rat heart λZAP cDNA library. The deduced primary structure of the protein encoded by the open reading frame corresponds to 971 amino acids, that can be organized into 12 transmembrane helices. The cloned gene was functionally expressed in HeLa cells. Maximal expression was detected 18 h after transfection, after which transport activity rapidly declined. The electrogenic properties of the cloned transporter were demonstrated following reconstitution of the expressed exchanger protein into a tightly sealed phospholipid membrane.