牙鲆肝脏抗菌肽-2基因序列和表达分析

doi:10.3969/j.issn.1000-7091.2012.z1.003

摘要/Abstract

摘要： 抗菌肽是鱼类先天免疫的组成部分，发现新的抗菌肽对于动物抗病机理的研究和寻找新药物有重要意义。经过对GenBank数据库中牙鲆EST文库进行同源比对后，筛选得到一个具有完整编码框的抗菌肽cDNA序列，结果再经过RT-PCR进行验证，命名为LEAP-2，对该序列进行注释，获得序列登录号EU586111。该抗菌肽能编码102个氨基酸的多肽，前28个氨基酸组成信号肽。推测编码蛋白质分子量为11 276.9 u，理论等电点(PI)为9.33。其蛋白质序列与另外9种鱼类的同源抗菌肽序列比较发现，同源性介于38.2%～61.7%。其中57至92位氨基酸为高度保守区，是维持其空间结构的主要区域，其中包含有4个保守的半胱氨酸，可能组成2个二硫键。荧光定量PCR分析显示，LEAP-2基因在肝脏中高表达，在牙鲆仔鱼不同发育时期中也有表达，但是表达水平存在明显差异，特别是在变态中期表达量比较高。

关键词: 牙鲆, 肝抗菌肽2, 序列分析, 同源建模

Abstract: Antimicrobial peptides are found in several kinds of tissues of many vertebrates and invertebrates． These peptides are predicted to operate as a first-line host defense mechanism exerting broad-spectrum activity against pathogenic bacteria， fungi， parasites， and enveloped viruses． Followed basic local alignment search to the flounder EST library of GenBank database by using LEAP-2 from Oncorhynchus as probe sequence， an cDNA sequence owned a complete open reading frame of antimicrobial peptides was achieved． The sequence was verified byRT-PCR and named LEAP-2， accession number of EU586111 was acquired after sequence annotation and submittingto the GenBank database． Flounder LEAP-2 gene encode a peptide of 102 amino acids with a 28 amino acids signalpeptide and a mature peptide of 74 amino acids， with a predicted mulecular weight of 11 276． 9 u and pI at 9． 33 ．The amino acid and nucleotide sequences were conserved between flounder and other organisms and the homologyranged 38． 2% － 61 ． 7% ， the 57 － 92 amino acids are highly conserved region is to maintain the spatial structure，including four conserved cysteine， may be composed of two disulfide bonds． The flounder LEAP-2 gene is expressedin a wide range of tissues and predominantly expressed in the liver． Its expression is developmentally regulated withno detection in early stages of metamorphosis but highly in the mid-metamorphosis．

Key words: Paralichthys olivaceus, Liver-expressed antimicrobial peptide 2, Sequence analysis, Homology modelling

中图分类号:

S917.4

CHEN Xiao-wu, SHI Zhi-yi, LI Qian, JIANG Shan. Sequence and Expression Analysis of LEAP-2 Gene from Paralichthys olivaceus[J]. ACTA AGRICULTURAE BOREALI-SINICA, doi: 10.3969/j.issn.1000-7091.2012.z1.003.

http://www.hbnxb.net/CN/Y2012/V27/IS1/12

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