Molecular cloning and expression of the heat shock protein 70 gene in the Kumamoto oyster Crassostrea sikamea |
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Authors: | Taisei Nagata Mamoru Sameshima Taku Uchikawa Natsumi Osafune Takeshi Kitano |
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Affiliation: | 1.Kumamoto Prefectural Fisheries Research Center,Kami-Amakusa,Kumamoto,Japan;2.Department of Biological Sciences, Graduate School of Science and Technology,Kumamoto University,Kumamoto,Japan |
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Abstract: | Episodes of summer mortality of the Kumamoto oyster Crassostrea sikamea are a major problem for its cultivation. Expression of the heat shock protein 70 (HSP70) is induced by various environmental stresses, including heat. We cloned and sequenced hsp70 complementary DNA from C. sikamea to investigate the relationship between hsp70 expression and heat tolerance in this oyster. Quantitative real-time polymerase chain reaction was performed using gill tissue dissected from oysters before and after heat shock for 1 h. The results showed hsp70 expression was faster and greater in oysters cultured at 20–22 °C than at 10–12 °C, and survival was lower among oysters cultured at 20–22 °C than at 10–12 °C. Moreover, heat tolerance was investigated by a 1-h pre-heat treatment, followed by exposure to heat shock conditions 5 days later. Survival was higher and hsp70 expression was notably lower in oysters that received the pre-heat treatment compared with those that did not. We conclude that a pre-heat treatment of only 1 h may be useful for inducing heat tolerance in C. sikamea, and that a low level of hsp70 expression after heat shock is an important index in selecting for high heat tolerance in these oysters. |
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