Biotechnology Bulletin ›› 2014, Vol. 0 ›› Issue (3): 137-145.

• Papers • Previous Articles     Next Articles

Cloning and Expression Analysis of Na+-K+-ATPase α in Red Tilapia(Oreochromis sp.)at Different Water Salinities

Wang Zhongliang1,2, Zhang Jiandong1,2, Huang Jiansheng1,2, Tang Baogui1,2, Zhou Hui1,2, Shi Gang1,2, Pan Chuanhao1,2, Wu Zaohe3,4, Chen Gang1,2   

  1. (1. Fisheries College, Guangdong Ocean University,Zhanjiang 524088;2. Key Laboratory of Aquaculture in South China Sea for Aquatic Economic Animal,Regular High Education Institute of Guangdong Province,Zhanjiang 524088;3.Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals,Zhanjiang 524088;4. Zhongkai University of Agriculture and Engineering,Guangzhou 510225)
  • Received:2013-09-24 Online:2014-03-29 Published:2014-03-31

Abstract: The full-length cDNA of red tilapia(Oreochromis sp.)Na +-K +-ATPase α was cloned using homological cloning and rapid amplification of cDNA ends(RACE)methods. Results showed that the full-length cDNA was 3 379 bp, containing a 5'untranslated region(5'-UTR)of 143 bp, a 3'-UTR of 164 bp and open reading frame of 3 072 bp bp encoding 1 023 amino acids with an estimated molecular weight of 112.5 kD and an estimated isoelectric point of 5.26. BLAST analysis revealed that the Na +-K +-ATPase α of red tilapia shared a high homology of 97%-99% with the corresponding proteins of other know species, respectively. Phylogenetic analysis showed that Na +-K +-ATPase α of red tilapia shared closest relationship with the corresponding proteins of Oreochromis mossambicus and Sarotherodon melanotheron, respectively. The expression pattern of Na +-K +-ATPase α at different water salinity was analyzed by fluorescent quantitative real-time PCR technology, and results showed that the expression of Na +-K +-ATPase α reached the highest level at 25, and fell to a relatively lower level at 32. In the same salinities, the expression of Na +-K +-ATPase α mRNA increased significantly(P<0.05)during the adaptation phase(12 h), and in regulatory phase(after 24 h)the amounts of Na +-K +-ATPase α mRNA fell to a relatively lower level, while the expression levels were still higher than that of fish in control group(P<0.05).

Key words: Red tilapia(Oreochromis sp .), Na+-K+-ATPase, α Rapid amplification of cDNA ends(RACE), Fluorescent quantitative real-time, PCR