短尾蝮蛇毒磷脂结合抗凝蛋白β亚基的基因克隆和序列分析  

GENE CLONE AND SEQUENCE ANALYSIS OF THE β SUBUNIT OF PHOSPHOLIPID-BINDING ANTICOAGULATION PROTEIN FROM AGKISTRODON HALYS BREVICAUDUS VENOM

作　　者：黎渊弘[1] 林文珍[2] 臧宁[3] 梁莹[4] 黎肇炎[3] 

机构地区：[1]广西医科大学第一附属医院药剂科,南宁530021 [2]广西医科大学生物化学与分子生物学教研室 [3]广西医科大学医学科学实验中心 [4]广西医科大学微生物学与免疫学教研室

出　　处：《广西医科大学学报》2010年第2期188-191,共4页Journal of Guangxi Medical University

基　　金：广西研究生教育创新计划项目资助(No.2006105981007D10)

摘　　要：目的:克隆短尾蝮蛇毒磷脂结合抗凝蛋白(PBAP)β亚基基因,并对该基因进行序列分析。方法:从短尾蝮蛇的毒腺中提取总RNA,设计引物并采用RT-PCR技术进行体外扩增PBAPβ亚基cDNA序列,将PCR扩增产物克隆至载体PMD-19T,通过PCR进行鉴定后,提取阳性克隆菌体质粒进行测序。结果:序列分析结果显示:PBAPβ亚基cDNA序列长度为441bp,编码框由146个氨基酸组成,其中包括由23个氨基酸的信号肽,以及123个氨基酸组成的成熟肽。结论:PBAPβ亚基基因及其氨基酸序列和已报道的一些蛇毒凝集素β亚基的基因以及氨基酸序列相比较,有88%～99%的同源性。Objective：To clone and analyze sequence of the β subunit of phospholipi d-binding anticoagulation protein （PBAP）from Agkistrodon halys Brevicaudus venom. Methods： The total RNA was extracted from the venom gland of snake Agkistrodon halys Brevicaudus. The cDNA sequences of the β subunit of PBAP were amplified by RT-PCR using primers and cloned into the pMD 19-T Vector. Plasmid DNAs obtained from positive clones were identified by means of PCR reaction. The cDNA sequences were determined with positive clones directly. Results：The sequence analysis showed that the cDNA sequence of the β subunit of PBAP which encoded an open reading frame （ORF） of 146 amino acids was 441 bp, also included a signal peptide of 23 amino acids, mature protein of 123 amino acids. Conclusion：The β subunit of PBAP was compared with the amino acids sequence for other Lectin of snake venom protein β subunit （such as Deinagkistrodon acutus, Trimeresurus stejnegeri）, their homology was 88 % -99 %.

关 键 词：蛇毒 克隆 序列分析 

分 类 号：R914.1[医药卫生—药物化学]