猪α(1,2)岩藻糖转移酶基因(FUT1)启动子区结构特征及多态性分析  

Structural Feature and Polymorphism Analysis of Promoter Region of Porcine(Sus scrofa) α-1,2-fucosyltransferase Gene(FUT1)

作　　者：刘颖[1] 吴森[1] 夏日炜 孙寿永[1] 朱国强[2] 吴圣龙[1] 包文斌[1] 

机构地区：[1]扬州大学动物科学与技术学院/江苏省动物遗传繁育与分子设计重点实验室,扬州225009 [2]扬州大学兽医学院,扬州225009

出　　处：《农业生物技术学报》2015年第7期917-922,共6页Journal of Agricultural Biotechnology

基　　金：江苏省高校自然科学研究重大项目(No.14KJA230003);国家自然科学基金(No.31472066;No.31372285和No.31172183);扬州大学大学生科技创新基金;江苏高校优势学科建设工程资助项目(PAPD)

摘　　要：α-(1,2)岩藻糖转移酶基因1(FUT1)是断奶仔猪F18大肠杆菌(Escherichia coli)抗性的重要候选基因。为探讨FUT1基因启动子区的分子结构特征和重要的变异位点,本研究基于前期猪(Sus scrofa)FUT1基因上游序列转录活性双荧光素酶报告基因的检测结果,对表现出启动子活性的FUT1基因上游非编码区-1150~50 bp区域Cp G岛、启动子和转录因子结合位点等结构特征进行生物信息学分析,并利用聚合酶链式反应-单链构象多态(polymerase chain reaction-single strand conformation polymorphism,PCR-SSCP)方法对该段序列在野猪和11个不同猪品种中的多态性进行分析。实验结果表明,FUT1基因上游启动子区序列存在3个Cp G岛(Cp G islands),分别位于-1104^-886 bp、-796^-619 bp和-316^-130 bp;启动子和转录因子结合位点的特征分析显示,该序列存在特异性蛋白-1(specificity protein 1,Sp1)、核转录因子-2(nuclear transcription factor 2,NRF-2)、E26特异性转录因子(E26 transformation-specific,c-Ets)和GATA结合蛋白1(GATA-binding protein 1,GATA-1)等转录因子结合位点;PCR-SSCP结果显示,FUT1基因的启动子区存在一个T(-726)C的突变位点,共检测到AA、AB和BB 3种基因型,分型后计算出各群体的基因型和等位基因频率,分析结果显示,梅山、荣昌、小梅山AA基因型频率很低,而野猪以及二花脸、枫泾、淮猪、香猪等中国地方猪品种中仅检测到BB和AB基因型,未检测到AA基因型。本研究结果进一步证实,中国地方猪种和引进猪种F18大肠杆菌抗性的遗传基础存在明显差异,这种差异可能与FUT1基因启动子区T(-726)C位点变异有关。研究结果为可稳定遗传的功能多态位点的筛选以及开展FUT1基因的启动子区调控机理研究奠定基础。α- 1,2- fucosyltransferase gene（FUT1） is the important candidate gene of piglets＇ resistance to Escherichia coli F18. This study aimed to probe into the structures and important mutation loci of FUT1 gene.Based on the previous detection results which implied the transcription activity of FUT1 gene upstream sequence by dual- luciferase reporter, we screened the- 1150 to 50 bp Cp G islands, the promoter and the transcription factor binding sites of the FUT1 gene upstream sequence which showed promoter activity. Then,in this study, FUT1 gene upstream sequence was analyzed by cloning and bioinformatics, and the polymorphism in wild boar and 11 different pig breeds were investigated by polymerase chain reaction-single strand conformation polymorphism（PCR- SSCP）. The results showed that there were 3 Cp G islands which located at-1104--886 bp,-796 --619 bp and-316--130 bp; the analysis of characteristics of the promoter and transcription factor binding sites showed there were Sp1, NRF-2, c-ETS and GATA-1 existing; the results of PCR- SSCP analysis showed that there existed only a T（-726）C point mutation and AA, AB and BB genotypes were detected. Difference of genotype distribution between domestic pigs and foreign was significant. Genotype AA was undetectable in the Chinese native pig breeds such as Erhualian pigs, Fengjing pigs, Huai pigs and Xiang pigs,, and frequency of genotype AA was low in other Chinese native pig breeds（0.012-0.058）, genotype BB was dominant in wild boar and Chinese native pig breeds except Xiang pigs; but the frequency of genotype AA was higher in 3 western commercial breeds（Duroc, Yorkshire and Landrace）and 50% Duroc descent Sutai pigs and genotype BB barely existed（0.021-0.112）. This study further confirmed that the genetic basis which resistance to E. coli F18 was significant differences between Chinese native pig breeds and western pig breeds, which might be related to the T（-726）C point mutation in FUT1 gene promoter. This result provides a theoretical foundat

关 键 词：猪 α(1 2)岩藻糖转移酶基因(FUT1) 启动子区 多态性分析 

分 类 号：S858[农业科学—临床兽医学]