Molecular Cloning, Characterization and Expression Analysis of Wap65 Gene from the Crucian Carp
Molecular Cloning, Characterization and Expression Analysis of Wap65 Gene from the Crucian Carp
Jun Cui, Xiaoxu Zhou, Zhicheng Wang, Derong Kong, Xuemei Qiu, Hongdi Wang and Xiuli Wang*
ABSTRACT
The Wap65 is a warm temperature acclimation-related protein, and it plays an important role in adapting temperature shock. But the research about Wap65 in crucian carp is very limited. In this study, the CDS of Wap65 was firstly cloned and characterized from crucian carp (Carassius carassius). This sequence is 1338bp that encodes a polypeptide of 445 amino acids. The calculated molecular weight of crucian carp Wap65 protein (CcWap65) is 50.8kDa, containing a signal peptide cleavage site between amino acids 18 and 19. The CcWap65 is a hydrophilic protein and no trans-membrane topological proteins. The SMART analysis revealed that CcWap65 contains three hemopexin-like repeats (E-value < 0.05). The crucian carp Wap65 was mainly expressed in liver, with limited expression observed in intestine, skeletal muscle and kidney. CcWap65 was significantly up-regulated in the liver of crucian carp after heat stress, suggesting that increase in Wap65 gene may be related to high water temperature stress and play important roles in high water temperature environment of crucian carp.
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