[Cloning and Ion Transportation of Cryptosporidium andersoni ATP-binding Cassette 1 Gene].
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
; 34(4): 326-30, 2016 Aug.
Article
en Zh
| MEDLINE
| ID: mdl-30148291
ABSTRACT
Objective:
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.Methods:
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.Results:
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 ï¼5.51 ± 0.51ï¼, ï¼1.98 ± 0.06ï¼, ï¼108.33 ± 1.33ï¼ and ï¼0.93 ± 0.03ï¼ mmol/L in the blank group; ï¼6.25 ± 0.70ï¼, ï¼1.90 ± 0.13ï¼, ï¼107.73 ± 1.79ï¼ and ï¼0.87 ± 0.05ï¼ mmol/L in the control group; and ï¼14.84 ± 0.90ï¼, ï¼3.40 ± 0.14ï¼, ï¼127.64 ± 1.49ï¼ and ï¼1.72 ± 0.20ï¼ mmol/L in the transfected group. The concentrations of K+, Ca2+, Na+ and Mg2+ in extracellular fluid were ï¼12.72 ± 0.83ï¼, ï¼3.72 ± 0.03ï¼, ï¼116.83 ± 1.04ï¼ and (2.02 ± 0.18) mmol/L in the blank group; ï¼10.11 ± 0.90ï¼, ï¼3.58 ± 0.06ï¼, ï¼115.89 ± 1.86ï¼ and (1.71 ± 0.41) mmol/L in the control group; and ï¼5.77 ± 0.21ï¼, ï¼1.29 ± 0.18ï¼, ï¼96.21 ± 1.19ï¼ 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.Conclusion:
CaABC1 participates in the transportation of K+, Ca2+ and Mg2+.
Buscar en Google
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Transfección
/
Cryptosporidium
Límite:
Animals
Idioma:
Zh
Revista:
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
Asunto de la revista:
PARASITOLOGIA
Año:
2016
Tipo del documento:
Article
Pais de publicación:
CHINA
/
CN
/
REPUBLIC OF CHINA