凡纳滨对虾半胱次磺酸脱羧酶基因的克隆表达
欧英杰 1庄俊钰 2许银叶3
作者信息
- 1. 广东省食品工业研究所广东广州 510308; 广东省食品工业公共实验室广东广州 510308
- 2. 广东省食品工业公共实验室广东广州 510308
- 3. 广东省食品工业研究所广东广州 510308
- 折叠
摘要
牛磺酸(Taurine)是一种具有多种生物活性的含硫非蛋白氨基酸，在神经系统发育、眼睛发育、渗透压调节等方面有重要功能，目前在饲料、饮料、医药等行业有重要应用。同时其也是鱼类等水产动物风味形成的呈味氨基酸。半胱次磺酸脱羧酶(cysteine sulfinate decarboxylase, CSD)是牛磺酸生物合成的限速酶，在牛磺酸的合成中有重要功能。本研究利用race PCR技术克隆到了凡纳滨对虾(Litopenaeus vannamei)的半胱次磺酸脱羧酶基因(L. vannamei CSD, LvCSD)的全长序列，对其编码的蛋白的特性进行了分析。同时构建原核表达载体对其进行表达，获得可溶表达的LvCSD，并利用镍柱对其进行纯化，获得较纯的重组LvCSD。另外还发现LvCSD在较盐度为15‰的养殖水体中的凡纳滨对虾的血淋巴细胞和肝胰腺细胞中转录水平较高。
Abstract
Taurine is a nonprotein amino acid that has a great variety of biological activity, such as nervous system development, eyes development, osmotic regulation and other aspects. And it with important applications in the feed industry, drink industry as well as pharmaceutical industry so far. At the same time, it also the delicious amino acid of fish and other aquatic animal. And Cysteine sulfinate decarboxylase is a rate limiting enzyme in the biosynthesis of taurine. In this study, race PCR technology has been used to clone the full-length of cysteine sulfinate decarboxylase of Litopenaeus vanname (LvCSD). And the characteristics of LvCSD were analyzed. At the same time, prokaryotic expression vector was constructed for expression of LvCSD, and then soluble LvCSD was purified by nickel column. It also found that the expression of LvCSD at a salinity of 15‰ was upregulated in haemocytes and hepatopancreas of L. vanname.
关键词
凡纳滨对虾/牛磺酸/半胱次磺酸脱羧酶/克隆/表达Key words
Litopenaeus vanname/taurine/cysteine sulfinate decarboxylase/clone/expression引用本文复制引用
出版年
2015
NETL
NSTL
维普
万方数据