海洋科学2017，Vol.41Issue(1)：83-95,13.DOI:10.11759//hykx20160112002
凡纳滨对虾碱性磷酸酶和酸性磷酸酶基因的克隆、表达及盐度应答效应
cDNA cloning and gene expressionin response to salinity of alkaline phosphatase and acid phosphatase from Litopenaeus vannamei
摘要
Abstract
In this study, alkaline phosphatase (ALP) and acid phosphatase (ACP) genes were cloned from Lito-penaeus vannamei by the rapid amplification of cDNA endsmethod.The full-length cDNA sequence of ALP was 1910 bp, encoding 536 amino acids, whereas the full-length cDNA sequence of ACP was 1544 bp, encoding 439 amino acids.A salinity regulation experiment was performed with three treatments: 31‰ (control), 16‰, and 3‰salt water.ALP and ACP gene expression was determined in tissues of the hepatopancreas, muscle, stomach, intes-tine and gills with real-time PCR after 45 days in saline. RT-PCR reveals that the ALP gene was detected in five tissues with the highest expression in the hepatopancreas(P<0.05) and the lowestin theintestine (P<0.05). The ACP gene was also detected in five tissues, and the gene expression in the hepatopancreas and gills was higher than that in the other tissues (P<0.05). After the salinity regulation experiment, the ALP and ACP genes presented different expression patterns among the various tissues. In the stomach and gill tissues, the higher expression level of the ALP gene occurred in the 31‰ saline group compared with the other two salinity groups (P<0.05); in the hepato-pancreas and intestinal tissues, the expression level of the ALP gene in the 16‰ saline group was significantly lower than that in the other two salinity groups (P<0.05); in muscle tissue, the expression level of the ALP gene was the highest in the 31‰ saline group (P<0.05) and the lowest in the 16‰ salinegroup (P<0.05). Whereas in the hepatopancreas, the expression level of the ACP gene in the 31‰ salinegroup was significantly lower than that in the other two salinity groups (P<0.05); in muscle and gill tissues, the expression level of the ACP gene in the 31‰salinegroup was significantly higher than that in the other two salinity groups (P<0.05); in the intestines, the ex-pression of the ACP gene was the highest in the 3‰ salinegroupand was significantly higher than that in the other two groups (P<0.05); in the stomach, the expression level of the ACP gene was not significantly different among the three groups. These results provide useful data for the molecular mechanism of Litopenaeus vannamei phosphatase metabolism under conditions of low salinity.关键词
凡纳滨对虾(Litopenaeus vannamei)/ALP基因/ACP基因/克隆表达/盐度Key words
Litopenaeus vannamei/alkaline phosphatase/acid phosphatase/cloning and expression/salinity分类
农业科技引用本文复制引用
张明明,王雷,王宝杰,刘梅,战文斌,蒋克勇..凡纳滨对虾碱性磷酸酶和酸性磷酸酶基因的克隆、表达及盐度应答效应[J].海洋科学,2017,41(1):83-95,13.基金项目
国家自然科学基金项目(41271547, 41401644) （41271547, 41401644）
中国科学院战略性先导科技专项(XDA05010400)National Natural Science Foundation of China, No.41271547, 41401644 （XDA05010400）
Strategic Priority Research Program of the Chinese Academy of Sciences, No.XDA05010400 （）
