猪BPI基因部分cSNPs检测及其对蛋白结构功能的影响

doi:10.11843/j.issn.0366-6964.2013.07.001

畜牧兽医学报 ›› 2013, Vol. 44 ›› Issue (7): 1000-1007.doi: 10.11843/j.issn.0366-6964.2013.07.001

猪BPI基因部分cSNPs检测及其对蛋白结构功能的影响

吴正常¹，王靖¹，朱世平¹，赵乔辉¹，刘璐¹，訾臣¹，吴圣龙^1,2*，包文斌^1,2*

（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. 江苏省现代种猪分子选育工程技术研究中心，常州 213149）

收稿日期:2012-12-24 出版日期:2013-07-23 发布日期:2013-07-23
通讯作者: 包文斌，博士，研究员，主要从事猪遗传育种研究，E-mail: wbbao@yzu.edu.cn；吴圣龙，博士，研究员，主要从事猪遗传育种研究，E-mail: slwu@yzu.edu.cn
作者简介:吴正常(1987-)，男，江苏南京人，硕士生，主要从事猪抗病育种研究，E-mail: 634468202@qq.com
基金资助:
江苏高校优势学科建设工程资助项目(PAPD)；国家自然科学基金(31172183)；江苏省产学研前瞻性联合研究项目(BY2012157)；江苏省科技支撑计划（农业）（BE2010371；BE2012330；苏州市科技支撑计划(SN201013;SN201133)）

The Partial cSNPs Detection of Pig BPI Gene and Its Influence on Protein Structure and Function

WU Zheng-chang¹, WANG Jing¹, ZHU Shi-ping¹, ZHAO Qiao-hui¹, LIU Lu¹, ZI Chen¹,WU Sheng-long^1,2*, BAO Wen-bin^1,2*

(1. Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design of JiangsuProvince, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009,China; 2. Jiangsu Engineering Research Centre for Molecular Breeding of Breeder Pig, Changzhou 213149, China)

Received:2012-12-24 Online:2013-07-23 Published:2013-07-23

摘要/Abstract

摘要：

本试验以98头苏太猪为研究对象，检测BPI基因部分编码区的单核苷酸多态性（SNPs），并预测分析碱基突变对BPI蛋白结构和功能的影响。利用PCR-SSCP和PCR-RFLP结合测序的方法检测BPI基因外显子1、4和10的SNPs，运用生物信息学比较分析原蛋白和突变型蛋白的理化性质和结构。结果表明，以BPI基因cDNA序列作为参照，BPI外显子1、4和10存在6个SNPs，3个同义突变（c.24A>G、c.54T>C和c.522C>T）和3个错义突变（c.433C>T(Arg145Trp)、c.1060A>G(Thr354Ala)和c.1151T>G(Leu384Arg)）。3种错义突变未改变BPI蛋白的理化性质、信号肽、跨膜区、糖基化位点以及二、三级结构，但是导致磷酸化位点的改变或存在其他潜在功能位点区域。Western blot试验发现，BPI蛋白的分子量为53 ku，苏太猪F18大肠杆菌抗性型猪的BPI蛋白表达量显著高于敏感型猪。猪BPI外显子4和10的碱基突变不同程度影响到蛋白功能，并可能对机体疾病的抗性/易感性产生影响。

Abstract:

This study was conducted to detect the single nucleotide polymorphism (SNPs) of BPI gene in partial coding region in ninety-eight Sutai pigs, and to predict the effect of mutation on BPI protein structure and function.PCR-SSCP/RFLP combined with sequencing were used to detect the SNPs in exon 1, exon 4 and exon 10 of BPI gene, and bioinformatics approach was used to predict the physicochemical properties and structures of original protein or mutant protein. Refer to the cDNA sequence of BPI gene, six SNPs existed in exon 1, exon 4 and exon 10 of BPI gene, with three synonymous mutations（c.24A>G , c.54T>C and c.522C＞T） and three missense mutations (c.433C>T(Arg145Trp), c.1060A>G(Thr354Ala) and c.1151T>G(Leu384Arg)). Three missense mutations had no influence on the physicochemical properties, signal peptide, transmembrane region, glycosylation sites, secondary structure and tertiary structure of the BPI protein，but led to the change of its phosphorylation sites or the existence of other potential function sites domain. The result of Western blot experiment indicated that there was an target band at 53 ku. It was obvious that expression of BPI protein in ETEC F18-resistant group was significant higher than that in susceptible group. The base mutations in exon 4 and exon 10 of pig BPI gene affect protein function in one way or another, which may have an influence on resistance/susceptibility of the pig disease.

中图分类号:

Q38
S813.3

吴正常，王靖，朱世平，赵乔辉，刘璐，訾臣，吴圣龙，包文斌. 猪BPI基因部分cSNPs检测及其对蛋白结构功能的影响[J]. 畜牧兽医学报, 2013, 44(7): 1000-1007.

WU Zheng-chang, WANG Jing, ZHU Shi-ping, ZHAO Qiao-hui, LIU Lu, ZI Chen,WU Sheng-long, BAO Wen-bin. The Partial cSNPs Detection of Pig BPI Gene and Its Influence on Protein Structure and Function[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(7): 1000-1007.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/ 10.11843/j.issn.0366-6964.2013.07.001

https://www.xmsyxb.com/CN/Y2013/V44/I7/1000

参考文献

［1］ FISHER C J, MARRA M N, PALARDY J E, et al. Human neutrophil bactericidal/permeability-increasing protein reduces mortality rate from endotoxin challenge: a placebo-controlled study ［J］. Crit Care Med, 1994, 22(4): 553-558.

［2］JAGIELLO P, KLEIN W, SCHULTZ H, et al. Association study of Wegener granulomatosis and the functionally relevant A645G polymorphism in the bactericidal/permeability increasing protein (BPI) gene［J］. Int J Immunogenet, 2005, 32(1): 3-6.

［3］KLEIN W, TROMM A, FOLWACZNY C, et al. A polymorphism of the bactericidal/permeability increasing protein (BPI) gene is associated with Crohn's disease［J］. J Clin Gastroenterol, 2005, 39(4): 282-283.

［4］HUBACEK J A, PITHA J, SKODOVA Z, et al. Polymorphisms in the lipopolysaccharide-binding protein and bactericidal/permeability-increasing protein in patients with myocardial infarction［J］. Clin Chem Lab Med, 2002, 40(11): 1097-1100.

［5］OOI C E, WEISS J, ELSBACH P, et al. A 25-kDa NH2-terminal fragment carries all the antibacterial activities of the human neutrophil 60-kDa bactericidal/permeability increasing protein［J］. J Biol Chem, 1987, 262(31): 14891-14894.

［6］CHRISTOPHER K T, THOMAS J S, SHI X W, et al. Genetic markers for improved disease resistance in animals (BPI) ［J］. United States, Kind Code: Al, Patent Application, 2004, 23: 49-80.

［7］朱璟, 潘章源, 訾臣, 等. 11个猪种群体BPI基因第10外显子HpaⅡ遗传变异分析［J］. 中国畜牧杂志, 2011, 47(23): 14-17.

［8］杨静, 李成云, 王云月, 等. 酿酒酵母分泌蛋白组的计算机分析［J］. 中国农业科学, 2005, 38(3): 516-522.

［9］DENTENER M A, FRANCOT G J, BUURMAN W A. Bactericidal/permeability-increasing protein, a lipopolysaccharide-specific protein on the surface of human peripheral blood monocytes［J］. J Infect Dis, 1996, 173(1): 252-255.

［10］MANNION B A, KALATZIS E S, WEISS J, et al. Preferential binding of the neutrophil cytoplasmic granule-derived bactericidal/permeability increasing protein to target bacteria. Implications and use as a means of purification ［J］. J Immunol, 1989, 142(8): 2807-2812.

［11］TAKAHASHI M, HORIUCHI Y, TEZUKA T. Presence of Bactericidal/permeability increasing protein in human and rat skin［J］. Exp Dermatol, 2004, 13(1): 55-60.

［12］BEAMER L J, CARROLL S F, EISENBERG D. Crystal structure of human BPI and two bound phospholipids at 2.4 angstrom resolution［J］. Science, 1997, 276(5320): 1861-1864.

［13］ZOR T, MAYR B M, DYSON H J, et al. Roles of phosphorylation and helix propensity in the binding of the KIX domain of CREB-binding protein by constitutive (c-Myb) and inducible (CREB) activators［J］. J Biol Chem, 2002, 277: 42241-42248.

［14］CALARESE D A, SCANLAN C N, ZWICK M B, et al. Antibody domain exchange is an immunological solution to carbohydrate cluster recognition［J］. Science, 2003, 300(5628): 2065-2071.

［15］朱立平. 蛋白质糖基化与B细胞免疫［J］. 上海免疫学杂志, 2001, 21(4): 193-194.

［16］HUBACEK J A, BUCHLER C, ASLANIDIS C, et al. The genomic organization of the genes for human lipopolysaccharide binding protein (LBP) and bactericidal permeability increasing protein (BPI) is highly conserved［J］. Biochem Bioph Res Co, 1997, 236(2): 427-430.

［17］IOVINE N M, ELSBACH P, WEISS J. An opsonic function of the neutrophil bactericidal/permeability increasing protein depends on both its N- and C-terminal domains［J］. Proc Natl Acad Sci USA, 1997, 94(20): 10973-10978.

［18］叶兰, 訾臣, 刘璐, 等. 仔猪BPI基因表达水平与大肠杆菌F18菌株感染的关系［J］. 遗传, 2011, 33(11): 1225-1230.

[1]	李想，鄢新义，曹露，田佳，秦春华，李委奇，温万，石万海，张胜利，张沅，王雅春. 新旧版本《中国荷斯坦牛体型鉴定规程》对比分析[J]. 畜牧兽医学报, 2016, 47(12): 2414-2419.
[2]	李建，宁静，陈西锐，宋成艳，雷安民. 小鼠H1foo基因四环素诱导表达系统的构建与表达[J]. 畜牧兽医学报, 2014, 45(5): 757-762.
[3]	左清清, 姚娜，董坤哲，叶绍辉，马月辉. 低氧适应的线粒体调控机制研究进展[J]. 畜牧兽医学报, 2013, 44(7): 993-999.
[4]	刘青青，郑丽明，刘红亮，苏建民，权富生，张涌. 小鼠早期胚胎发育过程中的DNA去甲基化[J]. 畜牧兽医学报, 2013, 44(4): 501-507.
[5]	齐超;谢岩;吴晓平;姜力;刘剑锋;孙东晓;张勤;张沅. 基于全基因组信息鉴定中国荷斯坦牛产奶性状基因及功能注释[J]. 畜牧兽医学报, 2012, 43(6): 872-877.

猪BPI基因部分cSNPs检测及其对蛋白结构功能的影响

The Partial cSNPs Detection of Pig BPI Gene and Its Influence on Protein Structure and Function

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 5

编辑推荐

Metrics

本文评价