阿拉善双峰驼ENaCα亚基基因克隆及激素对肾皮质细胞中该基因表达的影响

doi:10.16431/j.cnki.1671-7236.2019.02.002

Abstract

Abstract:

This study was aimed to explore the effect of sodium channel in epithelial cells of Alax League Bactrian camel.PCR primers were designed using the ENaCα subunit gene fragment which was provided by Shenzhen Huada company.Total RNA was extracted from the renal cortex of Alax League Bactrian camel,and cDNA was obtained by reverse transcription synthesis,then the renal cortex ENaCα subunit gene fragments were amplified by PCR.The cloning plasmid pMD19-T-ENaCα was constructed,the recombinant plasmid was identified by double enzyme digestion and squencing.The renal cortex of Alax League Bactrian camel was collected,after the cells were cultured in the P3 generation,different concentrations of aldosterone and angiotensin Ⅱ were added,the expression of ENaCα subunit gene was detected by Real-time quantitative PCR to determine the sensitivity of ENaCα changes in two hormone concentrations.The results showed that the ENaCα subunit gene was successfully cloned with the length of 2.2 kb,double digestion with Sma Ⅰ and Pst Ⅰ gave 2.2 and 2.7 kb bands,the recombinant plasmid pMD19-T-ENaCα was successfully constructed.Real-time quantitative PCR results confirmed that the expression levels of ENaCα treated with 1×10^-9 mol/L aldosterone and angiotensin Ⅱ were significantly higher than that of control group (P<0.05);The expression levels of ENaCα treated with 1×10^-6,1×10^-7 and 1×10^-8 mol/L aldosterone and angiotensin Ⅱ were significantly lower than that of control group (P<0.05).The results provided the experimental evidence for revealing the physiological mechanism of water and salt metabolism in Bactrian camel.

Key words: Alax League Bactrian camel; epithelial sodium channel; aldosterone; angiotensin Ⅱ

CLC Number:

S824

ZHU Chunxiao, SUN Rui, ZHANG Yanru, ZHANG Dong, LIU Chunxia, MENG Fanhua. Cloning of ENaCα Subunit Gene from Alax League Bactrian Camel and Effect of Hormone on the ENaCα Expression of Renal Cortex Cells[J]. , 2019, 46(2): 338-344.

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Cloning of ENaCα Subunit Gene from Alax League Bactrian Camel and Effect of Hormone on the ENaCα Expression of Renal Cortex Cells

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