渔业科学进展2015，Vol.36Issue(2)：63-70,8.DOI:10.11758/yykxjz.20150208
脊尾白虾(Exopalaemon carinicauda)α2-巨球蛋白cDNA全长的克隆和表达分析
The Cloning and Expression of Alpha2-Macrogloblin Gene ofExopalaemon carinicauda
摘要
Abstract
Exopalaemon carinicauda is one of most commercially important shrimps in China Sea due to its advantage in reproduction, longer growing seasons, and ecological flexibility. However, a variety of shrimp diseases have become an increasingly serious issue that lead to economic loss.E. carinicauda relies on the innate immunity to fight against invading pathogens. Alpha2-Macrogloblin (α2M) is an inhibitor of diverse proteases ubiquitous in animals. Alpha2M enfolds the target proteases and block their interactions. Thereforeα2M plays an important part in avoiding the deleterious effects of its active components. Based on the EST sequence from a cDNA library ofE. carinicaudahemocyte,we cloned the cDNA of alpha2-macrogloblin ofE. carinicauda(namelyEcα2M) using rapid amplification of cDNA ends (RACE). TheEcα2M cDNA was 4823 bp in length, which contained an open reading frame (ORF) of 4413 bp, a 64 bp 5' untranslated region, and a 346 bp 3' untranslated region. The open reading frame encoded an N-terminal signal sequence of 23 residues, and a mature protein consisting of 1470 amino acids. The molecular mass of this protein was 163.0 kDa and the estimated pI was 5.03. Homology analysis revealed that the amino acid sequence of Ecα2M was highly similar to its homologs in other crustaceans. The similarity of Ecα2M was 80% betweenE. carinicauda andMacrobrachium rosenbergii. We also analyzed the expression of Ecα2M in different tissues with quantitative real-time PCR. Ecα2M was detected in all tested tissues ofE. carinicauda, including hemocytes, gill, hepatopancreas, muscles, ovary, intestine, stomach and eyestalk, and the expression was the highest in hemocytes. After challenged withVibrio anguillarum and white spot syndrome virus (WSSV), the expression ofEcα2M was up-regulated in hemocytes and hepatopancreas ofE. carinicauda. Our results suggested that Ecα2M might play an important role in the prawn immune response.关键词
脊尾白虾/α2-巨球蛋白/基因克隆/基因表达Key words
Exopalaemon carinicauda/alpha2-macrogloblin/Gene cloning/Gene expression分类
农业科技引用本文复制引用
王有昆,刘萍,段亚飞,李吉涛,李健..脊尾白虾(Exopalaemon carinicauda)α2-巨球蛋白cDNA全长的克隆和表达分析[J].渔业科学进展,2015,36(2):63-70,8.基金项目
国家虾产业技术体系项目(CARS-47)、国家自然科学基金课题(31472275)和南通市科技项目(HL2013010)共同资助. （CARS-47）
