杂色鲍β-1,3-葡聚糖识别蛋白基因的克隆和表达  

Molecular cloning and characterization of beta-1, 3-glucan recognition protein from small abalone Haliotis diversicolor

作　　者：王宝珍[1] 张子平 王艺磊[1] 王国栋[1] 邹志华[1] 王淑红[1] 贾锡伟[1] 

机构地区：[1]集美大学水产学院水产科学技术与食品安全重点实验室,福建厦门361021 [2]美国德克萨斯州立大学化学与生化系,德克萨斯圣马克斯78666

出　　处：《海洋科学》2011年第8期67-75,共9页Marine Sciences

基　　金：国家自然科学基金资助项目(20877034);福建省自然科学基金资助项目(2009J0102);福建省教育厅A类项目(JA08137);集美大学创新团队基金资助项目(2010A001)

摘　　要：从杂色鲍(Haliotis disversicolor)的表达序列标签(EST)中首先鉴定出β-1,3-葡聚糖识别蛋白基因片段(beta-1,3-glucan recognition protein,saβgrp)。通过5′RACE获得5′端序列,并用由头至趾(head to toe)PCR方法检测全长cDNA序列的正确性。saβgrp的cDNA全长为1 459 bp,包括387 bp的5′非编码区,987 bp的开放阅读框,85 bp的3′非编码区。saβgrp开放阅读框编码328个氨基酸。与现有报道的β-1,3-葡聚糖识别蛋白均含信号肽不同,本研究所获得的杂色鲍saβGRP不含信号肽,为非分泌型蛋白。实时定量PCR分析表明:经副溶血弧菌诱导后,24 h的实验组saβgrp的表达水平极显著高于对照组(P<0.01),48 h的实验组saβgrp基因显著高于对照组(P<0.05)。The cDNA of the beta-l,3-glucan recognition protein in small abalone Haliotis diversicolor （saβgrp） was cloned. It was originally identified from an expressed sequence tag （EST） fragment in a normalized cDNA library. Its 5＇ cDNA end was obtained by 5＇ rapid amplification of cDNA end （RACE） techniques and its complete sequence was confirmed by head to toe PCR. The full length cDNA of saflgrp was of 1459 bp, consisting of a 5＇-terminal UTR of 387 bp, an open reading frame of 987 bp, and a 3＇-terminal UTR of 85 bp. The deduced protein was composed of 328 amino acids, with an estimated molecular weight of 36.9 kDa and a predicted pI of 5.90. Different from other reported GRPs that contain a signal peptide, the putative saI3GRP does not contain a signal peptide and thus belongs to non-secretary proteins. Real time quantitative PCR analysis revealed that after Vibrio Parahaemolyticus injection, the expression level of saβgrp in hepatopancreases at 24 h was most significantly higher than that of the control group （P〈0.01）, while at 48 hour post-injection, it was significantly higher than that of the control group （P〈0.05）.

关 键 词：saβgrp 副溶血弧菌 杂色鲍(Haliotis disversicolor) 

分 类 号：Q751[生物学—分子生物学]