Journal of Tropical Oceanography ›› 2010, Vol. 29 ›› Issue (5): 125-131.doi: 10.11978/j.issn.1009-5470.2010.05.125cstr: 32234.14.j.issn.1009-5470.2010.05.125

• Marine biology • Previous Articles     Next Articles

Molecular cloning and sequence analysis of pancreatic trypsinogen and amylase from orange-spotted grouper (Epinephelus coioides)

HU Yong-le1, LIANG Xu-fang1, WANG Lin1, LI Guan-gui1, LIU Xiu-xia1, WANG Yun-xin2, ZHANG Hai-fa2   

  1. 1. College of Life Science and Technology, Jinan University, Guangzhou 510632, China; 2. Guangdong Daya Bay Fisheries Devel-opment Center, Huizhou 516081, China
  • Received:2008-09-03 Revised:2009-03-30 Online:2010-10-30 Published:2011-10-10
  • Contact: 梁旭方。 E-mail:tliangxf@jnu.edu.cn
  • About author:胡永乐(1986—), 女, 江西省南丰县人, 硕士研究生, 主要从事鱼类代谢基因的研究。E-mail: hu.yongle@yahoo.com.cn
  • Supported by:

    国家科技部863项目(2007AA09Z437); 国家自然科学基金项目(30670367); 广东省科技计划项目(2007B020701002,
    2005B20301005); 广东省自然科学基金项目(031886); 教育部留学回国人员科研启动基金项目

Abstract:

Two full-length cDNA sequence encoding pancreatic trypsinogen (TRY) and amylase (AMY) genes were isolated from orange-spotted grouper (Epinephelus coioides) by RT-PCR and RACE methods. The sequences were deposited in the GenBank database under accession number EU715402 and EU715401. The cDNA of TRY was 911bp, encoding 242 amino acids. The deduced amino acid sequence of trypsinogen has a catalytic triad, and sequence analysis reveals that the identities of trypsinogen amino acid are 59.9%–89.7% between fish and mammals. The obtained AMY was 1657bp in length, encoding 512 amino acids. The deduced amino acid of amylase contains eight α-helical and eight β-sheet structures, and the identities of amylase amino acid among fish, mammals and bird are 70.1%–91.8%, which suggests that amylase is relatively more con-served. This research will help to further study the regulation of the two genes and to develop dietary additives to stimulate their expression levels.

Key words: trypsinogen, amylase, gene cloning, sequence analysis, Epinephelus coioides