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Rationale: The paper reports the cloning of the Na+/Ca2+ exchanger NCX3 and characterizes its transporter activity. Figure 6a reports an experiment where the NCX3 mRNA was injected in Xenopus oocytes (control- water injected). Na+ gradient-dependent labelled (45)Ca2+ uptake was measured under the following experimental conditions: -internal Na+ was elevated by incubation with nystatin. - the external solution contained labelled CaCl2 in a K+ medium, in the absence of external Na+. Figure 6b presents the results of a similar experiment but in BHK cells. The cells were transfected with the NCX3 DNA (control - non transfected cells). The specific Na+-dependent uptake of (45)Ca2+ is plotted. Under these conditions, the labeled Ca2+ uptake was larger than in the control. Background uptake levels were estimated by exchanging the external K+ ions for Na+, thereby eliminating the Na+ gradient which is necessary for Ca2+ uptake. Taken together, the experiment demonstrates that the identified cDNA codes for a Na+/Ca2+ exchanger.. Experimental description: A unique exchanger clone from a rat brain cDNA library was isolated. * 'Therefore, we subcloned a poly(A)1-containing fragment of DNA into the BglII-NotI site of pIII (Fig. 1). This replaced most of the original NCX3 39-UTR with the 39-UTR derived from the SGLT1 (Na1-glucose cotransporter) clone (Hediger et al., 1987)' as described previously (Li et al., 1994). The

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  • Rationale: The paper reports the cloning of the Na+/Ca2+ exchanger NCX3 and characterizes its transporter activity. Figure 6a reports an experiment where the NCX3 mRNA was injected in Xenopus oocytes (control- water injected). Na+ gradient-dependent labelled (45)Ca2+ uptake was measured under the following experimental conditions: -internal Na+ was elevated by incubation with nystatin. - the external solution contained labelled CaCl2 in a K+ medium, in the absence of external Na+. Figure 6b presents the results of a similar experiment but in BHK cells. The cells were transfected with the NCX3 DNA (control - non transfected cells). The specific Na+-dependent uptake of (45)Ca2+ is plotted. Under these conditions, the labeled Ca2+ uptake was larger than in the control. Background uptake levels were estimated by exchanging the external K+ ions for Na+, thereby eliminating the Na+ gradient which is necessary for Ca2+ uptake. Taken together, the experiment demonstrates that the identified cDNA codes for a Na+/Ca2+ exchanger.. Experimental description: A unique exchanger clone from a rat brain cDNA library was isolated. * 'Therefore, we subcloned a poly(A)1-containing fragment of DNA into the BglII-NotI site of pIII (Fig. 1). This replaced most of the original NCX3 39-UTR with the 39-UTR derived from the SGLT1 (Na1-glucose cotransporter) clone (Hediger et al., 1987)' as described previously (Li et al., 1994). The
Title
  • Slc8a3_MF_846
Date
  • 2017-08-25
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