引用本文:	杨顶珑,韦秀梅,杨建敏,杨嘉龙,徐洁,王圣,刘相全,张艳敏.大竹蛏(Solen grandis)铁蛋白基因的克隆及其在转录水平上对微生物多糖的应答.海洋与湖沼,2013,44(3):664-669.

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*大竹蛏(Solen grandis)铁蛋白基因的克隆及其在转录水平上对微生物多糖的应答*
杨顶珑^1,2, 韦秀梅², 杨建敏², 杨嘉龙³, 徐洁^1,2, 王圣^1,2, 刘相全², 张艳敏⁴
1.上海海洋大学水产与生命学院;2.山东省海洋水产研究所山东省海洋生态修复重点实验室;3.中国科学院烟台海岸带研究所;4.山东商务职业学院

摘要:

本研究克隆获得了一个大竹蛏(Solen grandis)铁蛋白(SgFer)基因的cDNA全长, 其序列全长为848bp, 5’和3’端的非编码区分别为111bp和212bp, 开放阅读框525bp, 推测编码174个氨基酸, 预测分子量为20.2kDa, 理论等电点为5.20。通过荧光定量PCR法研究了SgFer在健康大竹蛏组织中以及大竹蛏受到微生物多糖刺激后的表达规律, 结果表明, SgFer在血细胞、鳃、外套膜、肌肉、性腺和肝胰腺等组织器官中均有表达, 在肝胰腺中的表达量最高。SgFer在大竹蛏受到肽聚糖(PGN)和葡聚糖(glucan)刺激后, 其mRNA的表达量显著上调, 分别在刺激后6h和12h达到最高; 而脂多糖(LPS)刺激不能诱导其mRNA表达量上调。SgFer可能参与大竹蛏对微生物多糖的清除反应。

关键词: 大竹蛏铁蛋白基因克隆荧光定量PCR

DOI：10.11693/hyhz201303019019

分类号:

基金项目:山东省农业良种工程课题“优质高产抗逆贝类良种选育”, 2009—2013; 国家自然科学基金项目, 31202025 号; 水生动物营养与饲料“泰山学者”岗位经费资助

附件

CLONING OF A FERRITIN GENE FROM SOLEN GRANDIS AND ITS RESPONSE TO MICROORGANISM GLYCAN ON TRANSCRIPTION LEVEL

YANG Ding-Long^1,2, WEI Xiu-Mei², YANG Jian-Min², YANG Jia-Long³, XU Jie^1,2, WANG Sheng^1,2, LIU Xiang-Quan², ZHANG Yan-Min⁴

1.College of Fisheries and Life Science, Shanghai Ocean University;2.Shandong Provincial Key Laboratory of Restoration for Marine Ecology, Shandong Marine Fishers Research Institute;3.Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences;4.Shandong Business Institute

Abstract:

In this study, the full length cDNA of a ferritin gene from Solen grandis (designed as SgFer) was cloned, and it was 848bp and consisted of a 5′ untranslated region (UTR) of 111bp, a 3′ UTR of 212bp and an open reading frame (ORF) of 525bp encoding a polypeptide of 174 amino acids with an estimated molecular mass of 20.2kDa and isoelectric point of 5.20. The tissue and temporal expression of SgFer after microorganism glycan challenge was recorded by real-time PCR. SgFer transcript could be detected in all examined tissues with the highest expression level in hepatopancreas. The mRNA expression of SgFer was up-regulated after PGN or glucan stimulation, and reached peak at 6h and 12h post-challenge, respectively. No significant changes were observed after LPS stimulation. SgFer might be involved in elimination of microorganism glycan.

Key words: Solen grandis Ferritin Gene cloning Real-time PCR