罗氏沼虾Rab11基因cDNA克隆及其组织表达分析  

MOLECULAR CHARACTERIZATION AND EXPRESSION RESEARCH OF RAB11 FROM GIANT FRESHWATER PRAWN, MACROBRACHIUM ROSENBERGII

作　　者：夏正龙[1] 黄雪娜[1] 黄振远[1] 陈雪峰[1] 高强[1] 濮剑威[1] 慎佩晶[1] 彭菲[1] 杨国梁[1] 

机构地区：[1]浙江省淡水水产研究所,国家罗氏沼虾遗传育种中心,浙江省淡水水产遗传育种重点实验室,湖州313001

出　　处：《水生生物学报》2016年第3期443-450,共8页Acta Hydrobiologica Sinica

基　　金：浙江省自然科学基金(LY12C19008);国家科技支撑计划项目(2012BAD26B04);浙江省重大科技专项(2012C12907);浙江省科研院所专项(2014F50002)资助~~

摘　　要：为了发掘罗氏沼虾(Macrobrachium rosenbergii)免疫相关基因,研究Rab蛋白(Ras-related proteins in brain)在罗氏沼虾免疫应答中发挥的作用,研究采用RACE-PCR技术克隆了罗氏沼虾Rab11基因全长c DNA序列,记为Mr Rab11。全长1381 bp,包括226 bp的5′UTR,511 bp的3′UTR和645 bp的开放阅读框,编码214个氨基酸,含有一个Rab结构域。氨基酸序列比对显示,罗氏沼虾与冈比亚按蚊(Anopheles gambiae)、蚤状溞(Daphnia pulex)、叶蝉(Homalodisca vitripennis)Rab11一致性分别为82%、83%和82%。软件预测,Mr Rab11编码的蛋白分子量约为23.75 k D,等电点约为5.34。实时荧光定量表达分析表明,Mr Rab11基因在罗氏沼虾各组织中都有表达,肝胰腺中的表达量最高,其次是肌肉和肠道。在阴沟肠杆菌(Enterobacter cloacae)感染12h后,罗氏沼虾肝胰腺中Mr Rab11的表达量上升,显著高于对照组(P<0.05),推测这是Mr Rab11对阴沟肠杆菌的应激表达,Mr Rab11在肝胰腺中参与了罗氏沼虾免疫应答过程。Rabs play important role in vesicular transport in all eukaryotic cells. The current study successfully cloned the full-length cDNA of a Rab gene ofMacrobrachiurn rosenbergii （named MrRabl 1） by the rapid amplification of cDNA ends （RACE） techniques. The full-length ofMrRabl 1 cDNA was 1381 bp, including a 226 bp 5＇-terminal un- translated region （UTR）, a 511 bp 3＇UTR and a 645 bp open reading frame （ORF） encoding a polypeptide of 214 amino acids residues that contain a Rab domain. The calculated molecular mass of deduced MrRab 11 polypeptide was 23.75 kD and the isoelectric point was 5.34. BLASTX analysis revealed that MrRabl 1 had significant homology to Anopheles gambiae, Daphnia pulex and Homalodisca vitripennis with the identity of 82%, 83% and 82%, respectively. Quantita- tive real-time reverse transcription PCR analysis demonstrated a broad expression ofMrRabl 1 in many tissues with the highest level in hepatopancreas and then muscle and intestine. The expression level of MrRab 11 in hepatopancreas was significantly （P〈0.05） induced by Enterobacter cloacae at 12h. These results suggest that MrRabl 1 may play a role in the innate immunity of M. rosenbergii.

关 键 词：罗氏沼虾 Rab11 组织表达 免疫应答 

分 类 号：Q344.1[生物学—遗传学]