蚂蟥Guamerin基因的克隆及吸食后时空表达分析  

Cloning of Guamerin gene in Whitmania pigra and its spatio-temporal expression analysis after ingestion

作　　者：吴曼郡 郭巧生[1] 史红专[1] 刘飞[1] 吴斌[1] 施国伟[1] 闫士猛 勾玲[1] WU Man-jun;GUO Qiao-sheng;SHI Hong-zhuan;LIU Fei;WU Bin;SHI Guo-wei;YAN Shi-meng;GOU Ling(Institute of Chinese Medicinal Materials,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学中药材研究所

出　　处：《中国中药杂志》2018年第18期3605-3610,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81872960,81673538,81473306)

摘　　要：该研究对非吸血蛭蚂蟥体中弹性蛋白酶抑制剂基因(wp Guamerin)进行了克隆和生物信息学分析,同时对该基因在蚂蟥不同组织、不同吸食阶段的时空表达特点进行研究。结果表明,蚂蟥wp Guamerin基因全长295 bp,相对分子质量为9 145.95 Da,包含1个编码83个氨基酸的开放阅读框,与吸血蛭Guamerin氨基酸序列相似性均在29%~65%;wp Guamerin编码的蛋白无信号肽,属于亲水性蛋白,二级结构主要由α-螺旋、延伸链、折叠和无规则卷曲组成。wp Guamerin在肌肉中表达量显著高于唾液腺,在嗉囊和肠中无表达;吸食后wp Guamerin在肌肉和唾液腺的表达量分别在第1,3天达到最高。因此,wp Guamerin是有别于吸血蛭Guamerin的小分子多肽蛋白,在蚂蟥消化道中不表达,主要在肌肉中表达,吸食行为可刺激蚂蟥肌肉和唾液腺wp Guamerin基因的表达而不能诱导蚂蟥消化道wp Guamerin的表达。The present study cloned wp Guamerin gene from a non-bloodsucking leech( Whitmania pigra),and the bioinformatics analysis of the sequence was performed. Additionally,the effects of feeding duration on the expression profile of the wp Guamerin gene were explored. The results showed that its sequence consisted of 295 nucleotides encoding a peptide of 83 amino acids( Genbank:KX768545),and its relative molecular weight is 9 145. 95 Da. wp Guamerin does not encode proteins with a signal peptide,belonging to the hydrophilic protein. Its secondary structure is mainly composed of α-helix,extending chain,folding and random curl. Its similarity with other blood-sucking leeches ranges from 29% to 65%. The results revealed that wp Guamerin mRNA was detected higher expression in muscle than in salivary glands of Wh. pigra,and did not expressed in ingluvies and intestine. Its expression in muscle and salivary glands showed a single peak curve after feeding and the peak was observed in the 1 stand 3 rdafter feeding,respectively. In summary,wp Guamerin in Wh. pigra is a small molecule polypeptide protein and is different from the Guamerin in blood-sucking leeches. wp Guamerin does not express in the digestive tract of Wh. pigra,and mainly express in muscle. Feeding behavior would stimulate the expression of wp Guamerin gene in muscle and salivary glands,but not in digestive tract.

关 键 词：蚂蟥 非吸血蛭 弹性蛋白酶抑制剂 小分子多肽蛋白 吸食 

分 类 号：S865.4[农业科学—野生动物驯养]