猪Nramp1基因多态性研究进展

Nramp1(天然抗性相关巨噬蛋白)是一种离子转运蛋白,其蛋白可抵抗多种胞内寄生病虫杆菌侵染而发挥重要免疫功能,猪Nramp1基因蛋白多态性较高,且对仔猪抗病力影响较大。本文综述了猪Nramp1基因的结构、功能特点以及调节机制与疾病的相关性,并对猪Nramp1基因的发展进行展望,以期为家畜抗病育种提供理论依据。     

大肠杆菌刺激猪肠上皮细胞后Nramp1基因的表达变化分析

猪天然抗性相关巨噬细胞蛋白基因1(Naturalresistance-associated Marophage Protein 1,Nramp1)作为影响抗病力的重要候选基因之一,在机体的免疫过程中发挥着重要的调控作用。为了在细胞水平上探讨Nramp1基因与引起仔猪腹泻的致病性大肠杆菌的关系及其在大肠杆菌感染过程中发挥的免疫调控作用,本研究利用F18ab、F18ac和K88ac等3种致病性大肠杆菌刺激体外培养的猪肠上皮细胞IPEC-J2,并利用实时荧光定量PCR检测3种菌体刺激IPEC-J2后Nramp1基因的表达变化情况。结果表明:F18ab、F18ac和K88ac 3种大肠杆菌刺激IPEC-J2后,Nramp1基因的表达均发生极显著(P0.01)上调,差异倍数分别为10、8、11倍,且F18ab和K88ac 2种菌体刺激后Nramp1基因的表达上调程度均极显著高于F18ac。本研究结果提示,Nramp1基因的表达与大肠杆菌的侵染有很大的相关性,其在大肠杆菌侵袭猪肠道中发挥了重要的免疫调控作用。本研究在细胞水平分析探讨了Nramp1基因的表达与3种致病性大肠杆菌侵袭的关系,为Nramp1基因在大肠杆菌侵染机体的过程中发挥的免疫调控作用研究提供了参考依据。     

NRAMP1基因研究进展及其在抗病育种中的应用

抗病性能是当前畜禽育种的重要目标性状。NRAMP1蛋白可抵抗分枝杆菌、沙门氏菌等多种胞内寄生病原菌的侵染而发挥重要免疫功能，对畜禽机体抗病力影响较大。作者综述了NRAMP1基因的结构、功能、作用机制及其与抗病力的关系，并探讨了NRAMP1基因作为畜禽抗病候选基因及其应用于抗病力分子标记辅助选择的可行性。     

畜禽TLR4基因抗病研究进展

畜禽的抗病力作为实际育种与生产的重要目标性状之一,已引起科研人员和广大养殖户关注。TLR4基因作为革兰氏阴性菌细胞壁外膜成分中脂多糖(LPS)的主要受体,在畜禽机体免疫应答过程中发挥重要作用,对畜禽抗病力具有较大影响。主要叙述TLR4基因的作用、主要表达细胞、作用机理及其与畜禽抗病性的关系,并对TLR4基因作为畜禽抗病候选基因在实际生产育种中进行了展望。     

家兔组织器官中Nramp1基因表达差异研究

本研究旨在检测家兔Nramp1基因mRNA在不同组织中的表达水平,为进一步阐明家兔Nramp1基因的生物学功能提供有用的方法和信息。根据GenBank中家兔Nramp1基因序列设计引物,采用基于SYBR GreenⅠ的Real-time PCR对家兔心脏、肝脏、脾脏、肺脏、肾脏、胃、盲肠、回肠、淋巴结、肌肉、脂肪和大脑中Nramp1表达水平进行了定量分析。结果表明,Nramp1基因在肺脏组织中表达量最高,胃组织中最低,而在肌肉和盲肠组织中不表达。研究结果说明家兔Nramp1表达具有一定组织器官特异性。     

浦东白猪Nramp1基因第6内含子独特突变位点分析

Nramp1是学术界近年研究的猪重要抗病基因之一,其第6内含子作为Nramp1的分型重点片段,国内外的诸多猪种均有清晰的测序分析结果。本研究通过提取浦东白猪血液组织DNA,使用PCR技术分离和克隆Nramp1基因的第6内含子基因片段,测序后与以往文献中的其他猪种进行比较。结果发现,浦东白猪在该段基因中有3个特异的突变点普遍存在,分别是259 bp的A→G突变,331 bp的G→A突变和29 bp后1~2个T碱基的插入。这些突变的发现可以为浦东白猪的育种和Nramp1的抗病性研究提供遗传数据。     

迪庆藏猪Nramp1基因第二内含子和第二外显子多态性位点遗传分析

以迪庆藏猪为研究对象,通过测序技术对Nramp1基因内含子2和外显子2进行多态性位点遗传分析。结果发现:迪庆藏猪在Nramp1基因内含子2上存在6个SNP位点,在外显子2上存在1个SNP;各SNPs均处于Hardy-Weinberg平衡,多态信息含量和杂合度均小于0.5。通过该研究了解了迪庆藏猪Nramp1基因的变异情况及群体遗传特征,为下一步开展抗病毒基因的遗传育种奠定了理论基础。     

牛Nramp1基因5′调控区序列的克隆及序列分析

采用LA—PCR技术扩增牛Nramp1基因2515bp的5′调控区序列,构建了重组克隆载体pEASY—T3-Nramp1,对阳性克隆进行了PCR扩增、限制性酶切鉴定、DNA测序及生物信息学分析。结果表明,试验成功构建了包含Nramp1基因5′调控区的重组质粒。经同源性比对发现,Nramp1基因5′调控区在不同物种中具有一定的保守性,在转录起始位点近端的启动子区域,牛与人、鼠、猪、羊的同源性分别是60．50％,58．52％,72．18％,81．95％。经预测．该调控区富含GR、SP1、c—Ets-1、NF—W2等转录因子结合住点。本研究为进一步确定牛Nramp1基因核心启动子区域及该基因的表达调控奠定了理论基础。     

4猪种Nramp1基因第6内含子多态性研究

天然抗性巨噬蛋白(Nramp)基因是与人、鼠的一些病原微生物的易感性和抗性有关的重要候选基因。为了研究猪Nramp1基因的多态性,利用PCR-RFLP技术检测了杜洛克、大白猪、长白猪和合作猪共270头个体Nramp1基因第6内含子NdeⅠ酶切位点多态性。结果表明,4个猪种群共检测到3种基因型(AA、AB和BB),其中AB基因型为杜洛克、大白猪和长白猪的优势基因型;AA基因型为合作猪的优势基因型。经卡方适合性检测,杜洛克、合作猪和长白猪处于Hardy-Weinberg平衡状态(P>0.05),大白猪处于Hardy-Weinberg不平衡状态。多态信息含量分析显示,Nramp1基因的第6内含子NdeⅠ酶切位点在各猪种表现出中度多态性。     

荷斯坦牛免疫抗病相关Nramp1基因生物信息学分析

为研究牛的抗病能力,探究荷斯坦牛Nramp1基因所编码蛋白的结构与功能,本研究采用生物信息学分析的方法对荷斯坦牛Nramp1基因编码蛋白质的理化性质、疏水性/亲水性、信号肽跨膜区、跨膜结构域、N-糖基化位点和磷酸化位点、蛋白质二级结构、三级结构及蛋白质互作进行分析和预测.结果表明,荷斯坦牛Nramp1基因所编码蛋白全长...     

Genetic resistance to Salmonella infection in domestic animals

Genetic resistance to Salmonella infection in experimental animal models is well described. However, genetic resistance in domestic animals, which has potentially great value in terms of controlling Salmonella in the food chain, has been relatively poorly described. Recent advances in genetics and immunology have identified several factors that influence resistance in chickens and pigs in particular. Resistance to systemic salmonellosis in the chicken is encoded by a number of factors including Nramp1 (now termed Slc11a1) and a novel gene, SAL1 that leads to increased macrophage activity against Salmonella. Studies in outbred, and in particular, inbred chickens have revealed considerable differences in levels of colonization of the gastrointestinal tract and responses to vaccination. Factors influencing this appear to include innate immune function, MHC and Nramp. In pigs several immune factors, including polymorphonuclear cell activity, have been shown to influence resistance.     

Sequence analysis of the NRAMP1 genes from different bovine and buffalo breeds

The natural resistance associated macrophage protein 1 (Nramp1) has been reported to confer resistance or susceptibility to Mycobacterium bovis, Salmonella typhimurium, and Leishmania donovani in the mouse, Mus musculus. A Gly and Asp substitution at position 169 of the mouse Nramp protein is invariably associated with the resistant and susceptible phenotypes, respectively. The present study aimed to detect polymorphisms in the NRAMP1 gene from different cattle and buffalo breeds. Genomic DNAs from five breeds of cattle and four breeds of buffalo were used in the study. Sequencing showed two nucleotide substitutions found in intron 4, three in exon V, and ten in intron 5. An amino acid substitution was observed at nucleotide position 1202 in exon V of the Japanese black, Angus, Philippine and Bangladesh swamp-type buffaloes which coded for Thr, while the Korean cattle, Holstein, African N'dama, Indonesian swamp-type buffalo and the Bangladesh river-type buffalo had Ile. All the breeds of cattle and buffaloes tested in this study coded for Gly at the position in exon VI which corresponds to the same amino acid of the murine Nramp1-resistant phenotype at position 169. The phylogenetic relationship among the different breeds showed a cluster comprised mainly of cattle and another one mainly of buffaloes.     

Novel variants within the coding regions of the Slc11A1 gene identified in Bos taurus and Bos indicus breeds

Although in the cow the genetic resistance to brucellosis has been previously attributed to the Slc11A1 gene encoding Nramp1 protein, none of the mutations described to date seems to be the cause. To be able to associate another polymorphism of the gene to brucellosis resistance, we characterized the gene and identified in different breeds of Bos taurus and Bos indicus, six new variants among a total of 11 single nucleotide mutations, of which five occurred in the coding sequence (three are missense mutations), one in the promoter region and five in introns. The allelic and genotypic frequencies calculated revealed differences (p < 0.05) among the breeds studied.     

牛主要抗病候选基因的研究进展

牛抗病基因与其自身的抗病性和免疫能力有一定的相关性,因此研究相关抗病基因对提高牛自身免疫能力和选育优良后代有重要意义,文章着重介绍了国内外关于天然抗性相关巨噬细胞蛋白1（natual resistant macrophage protein 1,Nramp1）、主要组织相容性复合物（major histocompatibility complex,MHC）、甘露糖结合凝集素（maflnan binding lectin,MBL）、Toll样受体（Toll-like receptors,TLR）及干扰素基因（interferon,IFN）与牛某些疾病相关性的研究进展,及其是否可用于后代育种的分子标记,如MHC与奶牛的乳房炎密切相关;牛TLR4基因的外显子1760 C>T突变中的CC基因型可作为奶牛乳房炎抗性筛选中的分子标记等。笔者认为研究这些抗病基因可更深层次了解牛的抗病机制,同时借助先进的技术手段如分子标记、基因工程等,可更好的选育出抗病后代,抗病基因在育种中的应用将是未来研究抗病性和免疫能力的主要方向。     

Frequency of bovine Nramp1 (Slc11a1) alleles in Holstein and Zebu breeds

Natural resistance against brucellosis in cattle is linked to the Nramp1 gene, which encodes a divalent cation transporter that localizes in the phagolysosome membrane in macrophages. Nramp1 gene in mouse plays a critical role in innate immunity favoring bacterial killing by macrophages in addition to its influence on adaptative immunity. Polymorphisms at the bovine Nramp1 3' untranslated region (3'UTR), detectable by Single Strand Conformational Analysis (SSCA), are associated with natural resistance against brucellosis. Such polymorphisms are associated with variation in the number of GT repeats. This study compared the frequency of Nramp1 3'UTR polymorphisms between Zebu and European bovine breeds. Eighty-one Holsteins (Bos taurus taurus) and 167 Zebu (Bos taurus indicus), including the following breeds: Nelore (n=95), Guzerá (n=37), and Gir (n=35), totaling 248 pure breed cattle studied. DNA extraction was performed using the guanidium protocol and genotyping was performed by SSCA. DNA from cattle considered genotypically resistant to brucellosis resulted in a single band (homozygous) with 175bp, corresponding to the 3'UTR with 13 GT pairs (GT13), whereas DNA from genotypically susceptible cattle generated one single band with 177bp (homozygous GT14) or double bands with both 175 and 177bp, or 175 and 179bp (heterozygous GT13/GT14 or GT13/GT15, respectively). A marked difference in the frequency of alleles was detected between the Zebu and Holstein cattle. Holsteins had an extremely homogeneous genotype, with 100% of the individuals with a GT13 genotype. In sharp contrast the Nelore breed had the most heterogeneous genotype with four allelic combinations, namely, homozygous GT13, homozygous GT14, heterozygous GT13/GT14, and heterozygous GT13/GT15. When the Zebu breeds were compared to each other, the only significant difference observed was the frequencies of the genotypes GT13 and GT14 between the Nelore and Guzerá breeds. The knowledge of allelic frequencies in different breeds of cattle may prove to be very useful in the future for planning breeding strategies for selection of resistant cattle.     

猪抗病育种研究进展

猪病给养猪生产造成的经济损失约占总产值的12％～15％,提高猪抗病能力和抗病育种已成为目前研究的热点。作者综述了干扰素基因、NRAMP1基因、肠毒素大肠杆菌、MHC基因等候选基因的研究进展，以期为抗病育种提供借鉴。     

文山黄牛的现状与保种措施

文章阐述了文山黄牛的品种特性、现状和保种措施。文山牛保种可采用原产地活体保种,迁地活体保种和现代生物技术保种等多种方式。文山牛体型较大,性情温顺,产肉性能好、活动敏捷、结构紧凑、肌肉强健、肉品质优良、耐粗饲、适应性广、繁殖力和抗病力均较强等特点。文山牛是云南省一个地方良种,种质资源保护刻不容缓。