Cloning and differential expression of Na,K-ATPase in Penaeus vannameiby Hongyan Sun, Junde Dong, Chunhua Ren, Lvping Zhang, Xueming Dan, Chang Chen, Yuanzhou Zhang, Chaoqun Hu

Marine Biology Research

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Year
2015
DOI
10.1080/17451000.2015.1024131
Subject
Aquatic Science / Ecology, Evolution, Behavior and Systematics / Oceanography

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Cloning and differential expression of Na,K-ATPase in

Penaeus vannamei

Hongyan Sunabc, Junde Dongab, Chunhua Rena, Lvping Zhanga, Xueming Danc, Chang

Chena, Yuanzhou Zhanga & Chaoqun Hua a Key Laboratory of Tropical Marine Bio-resources and Ecology (LMB), Key Laboratory of

Applied Marine Biology of Guangdong Province & Chinese Academy of Sciences (LAMB),

South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou,

China b National Experiment Station of Tropical Marine Biology, Chinese Academy of Sciences,

Sanya, China c College of Animal Science, South China Agricultural University, Guangzhou, China

Published online: 29 Jul 2015.

To cite this article: Hongyan Sun, Junde Dong, Chunhua Ren, Lvping Zhang, Xueming Dan, Chang Chen, Yuanzhou Zhang & Chaoqun Hu (2015): Cloning and differential expression of Na,K-ATPase in Penaeus vannamei , Marine Biology Research,

DOI: 10.1080/17451000.2015.1024131

To link to this article: http://dx.doi.org/10.1080/17451000.2015.1024131

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ORIGINAL ARTICLE

Cloning and differential expression of Na,K-ATPase in Penaeus vannamei

HONGYAN SUN1,2,3, JUNDE DONG1,2, CHUNHUA REN1, LVPING ZHANG1,

XUEMING DAN3, CHANG CHEN1, YUANZHOU ZHANG1 & CHAOQUN HU1* 1Key Laboratory of Tropical Marine Bio-resources and Ecology (LMB), Key Laboratory of Applied Marine Biology of

Guangdong Province & Chinese Academy of Sciences (LAMB), South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, China, 2National Experiment Station of Tropical Marine Biology, Chinese Academy of Sciences,

Sanya, China, and 3College of Animal Science, South China Agricultural University, Guangzhou, China

Abstract

To better understand the adaptive strategies of osmotic and ionic regulation in Penaeus vannamei, a full-length cDNA sequence encoding the Na,K-ATPase alpha subunit was isolated and characterized by rapid amplification of the cDNA ends (RACE). The nucleotide sequence was 3953 bp in length and included a 300 bp 5? untranslated region (UTR), 536 bp 3?

UTR with an AATAA canonical polyadenylation signal sequence and poly (A) tail, and a 3117 bp open reading frame (ORF) encoding a 1038 amino acid protein. The calculated molecular weight of the protein was 115.4 kDa with an estimated pI of 5.29. This sequence was submitted to the National Center for Biotechnology Information (NCBI) GenBank under accession number KF765670. A phenetic analysis showed that the Na,K-ATPase ?-subunit of P. vannamei shared a high degree of identity with other species. The relevant mRNA was detected in all of the tissues examined by fluorescent quantitative real-time polymerase chain reaction. The expression level was higher in the gills and hepatopancreas, and lower in the guts, muscles and epithelia. Increased expression of Na,K-ATPase mRNA levels was observed in all of the tissues examined in response to salinity exposure, and there was a significant difference in the tissues of the gills and hepatopancreas (P < 0.05); however, no statistical difference was observed in the epithelia, guts and muscles compared with the controls (P > 0.05). The findings demonstrated that the Na,K-ATPase ?-subunit is highly conserved in crustaceans, and differential expression in different tissues of P. vannamei was observed, indicating that the gene might have multiple functions in the shrimp.

Key words: Differential expression, K-ATPase, molecular cloning, Na, Penaeus vannamei, RACE

Introduction

Penaeus vannamei Boone, 1931, which can be found on the Pacific coast of the Americas from northern

Mexico to Northern Peru in the wild, is a commercially important penaeid shrimp and is widely aquacultured (e.g. Bartlett et al. 2002). P. vannamei can tolerate a wide range of environmental salinities (0.5? 40 ppt; e.g. Bray et al. 1994; Roy et al. 2007), and its life cycle involves a variety of stages that occur in a number of different habitats and salinities. A well-developed osmoregulatory ability is crucial for

P. vannamei to survive in multiple environments (e.g. Menz & Blanke 1980; Li et al. 2010). It is currently one of the most important species of cultured shrimp worldwide, and much of the culture occurs at low salinity levels (Towle 1997; Furriel et al. 2000). Its euryhaline character is related to its possession of numerous ion-exchanger proteins, which enable adaptation to multiple environments.