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[Cloning and Ion Transportation of Cryptosporidium andersoni ATP-binding Cassette 1 Gene].

Authors: Ju-hua, Wang; Pan, Wang; Chuan-huan, Liu; Xiu-mei, Sun; Qian-qian, Xie; Xiu-heng, Xue;

[Cloning and Ion Transportation of Cryptosporidium andersoni ATP-binding Cassette 1 Gene].

Abstract

To investigate the transportation of intracellular and extracellular K(+), Ca(2+), Na(+) and Mg(2+) under the function of Cryptosporidium andersoni ATP-binding cassette (CaABC) 1 gene.CaABC1 gene was amplified by PCR using specifically designed primers. The eukaryotic expression plasmid pEGFP-C1-CaABC1 was constructed, and transfected into mouse intestinal epithelial cells via liposome transfection. The blank (with no transfection) and control groups (transfected with empty plasmid pEGFP-C1) were also set. Changes in intracellular and extracellular K(+), Ca(2+), Na(+) and Mg(2+) concentrations were examined by the ion concentration assay kit.PCR amplification resulted in a 544 bp product. The recombinant plasmid pEGFP-C1-CaABC1 was successfully constructed. Green fluorescence was seen in the control and transfected groups, but not in the blank group. The concentrations of K(+), Ca(2+), Na(+) and Mg(2+) in intracellular fluid were (5.51 ± 0.51), (1.98 ± 0.06), (108.33 ± 1.33) and (0.93 ± 0.03) mmol/L in the blank group; (6.25 ± 0.70), (1.90 ± 0.13), (107.73 ± 1.79) and (0.87 ± 0.05) mmol/L in the control group; and (14.84 ± 0.90), (3.40 ± 0.14), (127.64 ± 1.49) and (1.72 ± 0.20) mmol/L in the transfected group. The concentrations of K+, Ca2+, Na+ and Mg2+ in extracellular fluid were (12.72 ± 0.83), (3.72 ± 0.03), (116.83 ± 1.04) and (2.02 ± 0.18) mmol/L in the blank group; (10.11 ± 0.90), (3.58 ± 0.06), (115.89 ± 1.86) and (1.71 ± 0.41) mmol/L in the control group; and (5.77 ± 0.21), (1.29 ± 0.18), (96.21 ± 1.19) and (0.64 ± 0.02) mmol/L in the transfected group. There were significant differences in K+, Ca2+ and Mg2+ concentrations between the transfected group and the control group.CaABC1 participates in the transportation of K+, Ca2+ and Mg2+.

Keywords

Cytoplasm, Mice, Adenosine Triphosphate, Animals, Cryptosporidium, Epithelial Cells, Transfection, ATP Binding Cassette Transporter 1, Plasmids

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
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