| 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)。 |
| 关键词: saβgrp 副溶血弧菌 杂色鲍(Haliotis disversicolor) |
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| 基金项目: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) |
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| Molecular cloning and characterization of beta-1, 3-glucan recognition protein from small abalone Haliotis disversicolor |
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| Abstract: |
| The cDNA of the beta-1,3-glucan recognition protein in small abalone Haliotis disversicolor (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 saβgrp 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 saβGRP 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). |
| Key words: saβgrp Vibrio parahaemolyticus Haliotis disversicolor |
