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Title: Expression of MRNA and Protein of Na+-K+-ATPase Alpha Subunit in Gills of Tilapia (Oreochromis Mossambicus).
Authors: Hwang, P. P.;Fang, M. J.;Tsai, Jong-Chang;Huang, C. J.;Chen, S. T.
Contributors: 應用運動科學所
Keywords: tilapia;Na+-K+-ATPase a subunit;gills;cloning, Mrna;protein expression;antibody
Date: 1998-06
Issue Date: 2012-04-20T02:21:03Z
Publisher: Springer Verlag
Abstract: Using RT-PCR, partial cDNA of Na+-K+-ATPase a subunit of tilapia (Oreochromis mossambicus) was
cloned and sequenced. Clone TG3, with 1685 bp encoding a protein of 565 amino acids, showed higher
than 85% identity in deduced amino acid sequence with previously published animal Na+-K+-ATPase a-
1 subunit genes. Northern blot showed that TG3 is expressed in gills, kidneys and other organs in tilapia.
The amount of mRNA in gill tissue increased with the level of environmental salinity. A fragment of TG3
was constructed into a pQE30 expression vector, and the expressed fusion protein was used to raise the
antibody, Ab-TG3, in rabbit. Ab-TG3 was found not only to immuno-react with a major protein of
approximately 100 kDa in size in gills, kidneys, heart, and brain of tilapia, but also to recognize a similar
protein in gills of mudskipper (Periophthalmus cantonesis) and the kidneys, heart and brain of rat. Results
of immunoblotting indicate that the amount of Na+-K+-ATPase a submit in gills of seawater-adapted
tilapia was significantly higher than that in freshwater-adapted ones. Our results indicate that the salinitydependent
stimulation of mRNA of gill Na+-K+-ATPase a subunit is associated with corresponding
stimulation at the protein level. This provides direct evidence of enhanced transcription and translation
of Na+-K+-ATPase a-subunit gene upon salinity challenge.
Relation: Fish Physiology & Biochemistry, 18(4): 363-373
Appears in Collections:[應用運動科學研究所] 期刊論文

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