The Pig PeptideAtlas: A resource for systems biology in animal production and biomedicine

Biological research of Sus scrofa, the domestic pig, is of immediate relevance for food production sciences, and for developing pig as a model organism for human biomedical research. Publicly available data repositories play a fundamental role for all biological sciences, and protein data repositories are in particular essential for the successful development of new proteomic methods. Cumulative proteome data repositories, including the PeptideAtlas, provide the means for targeted proteomics, system‐wide observations, and cross‐species observational studies, but pigs have so far been underrepresented in existing repositories. We here present a significantly improved build of the Pig PeptideAtlas, which includes pig proteome data from 25 tissues and three body fluid types mapped to 7139 canonical proteins. The content of the Pig PeptideAtlas reflects actively ongoing research within the veterinary proteomics domain, and this article demonstrates how the expression of isoform‐unique peptides can be observed across distinct tissues and body fluids. The Pig PeptideAtlas is a unique resource for use in animal proteome research, particularly biomarker discovery and for preliminary design of SRM assays, which are equally important for progress in research that supports farm animal production and veterinary health, as for developing pig models with relevance to human health research.     

Genetically Modified Pig Models for Human Diseases

Genetically modified animal models are important for understanding the pathogenesis of human disease and developing therapeutic strategies.Although genetically modified mice have been widely used to model human diseases,some of these mouse models do not replicate important disease symptoms or pathology.Pigs are more similar to humans than mice in anatomy,physiology,and genome. Thus,pigs are considered to be better animal models to mimic some human diseases.This review describes genetically modified pigs that have been used to model various diseases including neurological,cardiovascular,and diabetic disorders.We also discuss the development in gene modification technology that can facilitate the generation of transgenic pig models for human diseases.     

筛选适用于小型猪遗传检测的微卫星位点

目的筛选用于封闭群小型猪遗传检测的微卫星位点。方法从资料和GenBank中选取扩增效果好、等位基因多、均匀分布于小型猪18条常染色体和X染色体上的100个微卫星位点,合成引物,对封闭群小型猪基因组进行PCR扩增及条件优化。PCR产物采用琼脂糖凝胶电泳、聚丙烯酰胺凝胶电泳和STR扫描技术进行分析和比较,选择多态性好的位点。结果筛选出32个分布于不同染色体且等位基因多的微卫星位点。结论筛选出了应用于封闭群小型猪遗传检测的微卫星位点。     

猪线粒体控制区微卫星(mtMs)变异及其分布特征

猪线粒体DNA长度因D-loop中mtMs序列重复数变异而不同。为阐明猪mtMs变异及其在不同群体中的分布特征,本研究对4个藏猪群体、八眉猪、烟台黑猪及3个引入猪群体共164个个体mtDNA D-loop全序进行测定,并与GenBank中发表的相关序列进行比对,分析了其中的重复单元核苷酸变异、重复次数及其在各群体间的分布规律。结果表明,因重复单元“5’-TGCGTACACG-3’”第2~4和10位上碱基变异,猪mtMs形成了以其为核心序列的多种重复单元(R^A^R^G)组成的复合结构,重复数介于3~47之间。单一重复(R^A)组成的mtMs结构在各群体中表现出分布优势,而大多数复合结构(R^AR^B)分布在国外选育群体中。藏猪的mtMs复合结构多达8个,R^AR^CR^E为其特有,另有5个与八眉猪共享。中国野猪和韩国地方猪种也有其特有mtMs结构。丰富的mtMs变异和特有结构与长期适应进化有关,本研究为地方猪种遗传资源及适应性研究提供了标记工具和理论基础。     

猪毛质量对胱氨酸提取率的影响

本文介绍了用不同方法处理的猪毛分别在实验室和工厂提取胱氨酸,证明胱氨酸的得率和猪毛质量有关。含杂１５％左右的干打猪毛大生产胱氨酸的得率平均４．４％;不含杂质的猪座子毛胱氨酸得率平均达６．７１％探讨了清除猪毛杂质,提高猪毛质量对胱氨酸生产的重要意义。     

Purification and Characterization of Four NADPH-Dependent Aldehyde Reductases from Pig Brain

By a procedure involving ammonium sulfate precipitation, gel filtration, and affinity chromatography, four aldehyde reductases (ALRs) were purified to enzymatic homogeneity from pig brain. These enzymes, designated ALR1, ALR2, ALR3, and succinic semialdehyde reductase were chemically and physically identical with, respectively, the high-Km aldehyde reductase, the low-Km aldehyde reductase, carbonyl reductase, and succinic semialdehyde reductase of other tissues and species. The purification procedure allows the purification of these enzymes from the same tissue homogenate in amounts sufficient for characterization and other enzymatic studies. This methodology should be applicable to the simultaneous and rapid purification of aldehyde reductases from other tissues.     

猪线粒体DNA多态性与中国地方猪种起源分化的关系

用24种限制性内切酶分析了我国21个具代表性地方猪品种、1个引进品种和2个来自中国和越南的野生近缘种mtDNA的RFLP。结果表明:在74个个体中检出的32种限制性态型可归结成7种单倍型,其间的差异主要来源于少数几个限制性位点的点突变;地方猪种4种单倍型间的平均遗传距离为0.143％,遗传多态程度(π值)仅为0.007％,说明遗传多样性非常贫乏,提示中国地方猪种可能起源于一个野猪亚种。     

猪Pit-1基因第三内含子序列的克隆测序

根据不同物种间同一基因核苷酸序列的保守性及相似性的特点 ,在人和鼠的Pit 1基因第三外显子上设计上游引物 ,而将下游引物设计在猪Pit 1基因第四外显子上。利用聚合酶链式反应 (PCR)技术 ,扩增出猪Pit 1基因第三内含子序列 ,并经由酶切及序列同源性比对确认该序列即为猪Pit 1基因第三内含子序列。此序列的确定为下一步进行遗传变异分析的研究奠定了基础     

猪脑组织总神经节苷脂提取纯化工艺的研究

本文较详细地研究论述了神经节苷脂（Ｇｌｓ）的提取纯化工艺。从猪脑组织中用萃取法获得Ｇｌｓ后,用多孔型弱碱性离子交换树脂３３５和强碱性离子交换树脂７１７对其进行分离纯化,并对工艺过程及产品品质进行了分析和比较研究,实验证明：３３５离子交换层析分离Ｇｌｓ,得率６５．９ｍｇ／１００ｇ湿组织,纯度８０．３％;７１７离子交换层析分离Ｇｌｓ,得率７７．４ｍｇ／１００ｇ湿组织,纯度７６．９％;从工艺过程及产品品质而言,采用多孔型弱碱性离子交换树脂分离提取Ｇｌｓ,具有一定的经济价值。     

Investigation of Tissue-Specific Human Orthologous Alternative Splice Events in Pig

Alternative splicing of pre-mRNA can contribute to differences between tissues or cells either by regulating gene expression or creating proteins with various functions encoded by one gene. The number of investigated alternative splice events in pig has so far been limited. In this study we have investigated alternative splice events detected in humans, in orthologous pig genes. A total of 17 genes with predicted exon skipping events were selected for further studies. The splice events for the selected genes were experimentally verified using real-time quantitative PCR analysis (qPCR) with splice-specific primers in 19 different tissues. The same splice variants as reported in humans were detected in 15 orthologous pig genes, however, the expression pattern predicted in the in silico analyses was only experimentally verified in a few cases. The results support the findings that splice events resulting in preservation of open reading frame are indicative of a functional significance of the splice variants of the gene.     

The Health of Ecology and the Ecology of Health

The health of ecology refers to ecosystem health—an extension of the concept of health to the ecosystem level. Health is reflected in the absence of distress syndrome, and by productivity, organization and resilience that characterize sustainability in the Earth's ecosystems. Transformation of ecosystems under stress from healthy to pathological conditions is often irreversible, even when the initial stress factors are removed. The ecology of health refers to the fact that human health is influenced by ecological conditions. The breakdown of ecosystems under stress are often conducive to an increase in human pathogens, recycling toxic substances, reduced yields and compromised food supplies, scarcity of potable water, and air pollution, all of which increases human health vulnerability. Addressing human health issues from an ecological perspective takes account of the social, ecological, and biophysical determinants. This opens the door to potential interventions “upstream”, in order to prevent illness, in addition to treating the disease once the malady has occurred.     

五指山猪生物学特性、易地繁育及遗传多样性研究

五指山猪是我国濒临灭绝的珍稀品种,为保护和利用这一特有种猪基因资源,我们开展了易地繁育和生物学特性及遗传多样性的研究。通过对其生长发育规律、繁殖生理、血液生理生化、遗传标记、DNA指纹图谱等项的观察和测试,发现五指山猪具有体型小、性成熟早、耐近交繁殖、遗传性较稳定、肉质好等特点。白细胞分类中淋巴细胞占77％,白细胞抗原(SLA) 血清学分型中,与其它品种之间存在有差异性。 用人源小卫星探针33.6和鸡小卫星探针cMS18,对五指山猪及长白猪和枫泾猪进行DNA指纹图比较分析,发现五指山猪的DNA指纹图相似系数,大大高于其它已研究过的正常体型猪种,说明它经历过较高程度的近交;以猪的生长激素（GH）cDNA为探针,用EcoRI、EcoRV、PstI等酶切基因组DNA进行RFLP分析时,五指山猪均出现了一条深色带,而长白猪和枫泾猪未发现。表明五指山猪在生长激素位点上与正常体型猪种亦存在差异。     

Analysis of antimicrobial peptides from porcine neutrophils

Cationic host defence peptides are important components of innate immunity in pigs and other mammalians. Most of these peptides have a direct antimicrobial activity and they also have a broad spectrum of effects on the host immune system, which may be taken into account in the introduction of novel therapeutics. Our method permits simultaneous isolation of six antibacterial peptides, i.e. prophenin-1, prophenin-2, PR-39, and protegrins 1-3 from a porcine neutrophil crude extract and characterisation of them. Among the obtained peptides the greatest bactericidal activity expressed as MBC was seen in protegrins (10 μg/ml), whereas in the other studied peptides MBC was on the level of 20 μg/ml. Minimal inhibitory concentrations (MIC) reached 10 μg/ml for protegrins 1-3 and 20 μg/ml for prophenins, and PR-39. Within the bactericidal range all isolated peptides didn't show cytotoxicity on cell lines used in our experiment.     

巴马香猪Toll样受体4基因cDNA的克隆及生物信息学分析

目的研究TLR4在猪自然免疫中的作用及机制,为抗病育种及免疫佐剂的开发提供依据。方法利用NCBI公布的TLR4基因序列设计引物,RT-PCR技术克隆巴马香猪TLR4基因。结果所得基因序列提交GenBank,登录号：GQ304754。经序列分析,发现巴马香猪TLR4基因开放阅读框长2526 bp,编码785个氨基酸,该蛋白等电点为6.58,分子量为96.4×103;与普通猪比对发现巴马香猪TLR4基因有5个碱基发生突变;与小鼠、狗、鸡、牛、羊和人的同源性分别为71.9%、81.5%、54.2%、86.4%、85.5%和81.9%;TLR4膜外区蛋白为背侧多个α螺旋和内侧多个β折叠平行交替排列构成一个弯曲状螺旋结构;N末端存在信号肽,且可能在23～24位氨基酸处存在裂解位点;胞外区有13个明显的LRR,分别位于第53～74、77～100、101～124、149～173、174～192、201～225、372～393、398～429、446～469、470～494、495～518、519～541、543～566位氨基酸区;膜外区含8个N连接的糖基化位点。结论本研究成功克隆巴马香猪TLR4基因,为进一步研究该基因的功能和蛋白质的特性奠定了基础。     

猪3号染色体九个微卫星位点的遗传多态性研究和遗传图谱构建

从已公布的猪 3号染色体连锁图谱中选取 9个微卫星位点 ,分析了这些位点在大白猪×梅山猪资源家系F2代 140头个体上的多态性 ,利用Crimap软件分别构建了猪 3号染色体两性平均连锁图谱以及公、母畜连锁图谱。结果表明 :各位点等位基因数为 2～ 4个 ,杂合度为 0 .436～ 0 .6 5 6 ,多态信息含量为 0 .35 1～ 0 .5 82 ;本研究所构建的平均连锁图谱全长为 16 1.1cM ,公、母畜连锁图谱全长分别为 135 .8cM和 188.7cM。与USDA所公布的连锁图谱相比 ,两者的标记顺序一致 ,但我们的图谱标记间隔偏大。     

Relationship Between Immune Cell Phenotypes and Pig Growth in a Commercial Farm

The objective of this study was to evaluate the relationship between the level and function of circulating immune cells with average daily gain, live and carcass measurements, feed intake, and feed conversion. Production performance was monitored throughout the pig's lifetime. Pigs were moved in weekly batches through the nursery and growing/finishing rooms at specific target weights. Animals were individually weighed at birth and at weaning, and then every two weeks while they were “on test” until they were “off test” and sent to the slaughterhouse. At six to seven weeks of age, the pigs were bled in the nursery. The percentage of immune cell subsets and lymphocyte proliferation was estimated using swine monoclonal antibodies and flow cytometric analysis. The predictive effect of the immune cell subset markers and lymphocyte proliferation on production traits was statistically analyzed. The results indicated that the proportion of several peripheral cell subsets, including CD16+, CD2+/CD16+, and CD8+ lymphocytes, appear to predict growth during the entire productive life of the pig. Larger percentages of lymphocytes expressing CD16+ CD2+/CD16+, and CD8+receptors in blood resulted in a reduction in average daily gain. In addition, high percentages of SLA-DQ+ cells were associated with better carcass weight and feed conversion. The CD16+, CD2+/CD16+, CD8+, and SLA-DQ± cell subsets appear to be important biomarkers involved with the inherent ability of the pig to efficiently grow and produce better carcass weight in representative commercial environments.     

猪心脏移植中供体急性脑死亡模型的初步研究

目的建立猪心脏移植供体的急性脑死亡模型,观察急性脑死亡前及后1、5、10 min时的血流动力学和血浆中儿茶酚胺释放的变化并进行初步探讨。方法采用30~40 kg的猪8只,急性脑死亡前为对照组,急性脑死亡后为实验组。测定脑死亡前基础水平及脑死亡后1、5、10 min时血流动力学改变和血浆中儿茶酚胺释放的变化,并对二者的变化进行初步探讨。结果急性脑死亡后1 min时血流动力学及儿茶酚胺水平改变最明显,心率增加了88%,收缩压升高了132%,心排量增加了80%,肾上腺素(E)和去甲肾上腺素(NE)分别升高240%和241%,多巴胺(DA)没有明显增加。随后二者均持续下降,至10 min时已降至基础水平以下。血流动力学改变的程度及时间分布与儿茶酚胺中的肾上腺素和去甲肾上腺素一致。结论急性脑死亡可造成机体血流动力学及血浆儿茶酚胺水平的剧烈改变,血中儿茶酚胺含量升高是造成急性脑死亡后血流动力学改变的原因,其中E和NE与血流动力学改变直接相关。     

The Fat Mass and Obesity-Associated (FTO) Gene is Associated with Intramuscular Fat Content and Growth Rate in the Pig

The association of the FTO gene with obesity has been implicated in various human populations. The FTO gene is also most likely involved in the regulation of energy balance and feed intake. Here, the FTO gene was studied as a candidate gene for fatness and growth rate traits in pigs. The amino acid sequence of the FTO gene showed high conservation among human, pig, and other important domestic animals. Twelve variants including ten SNPs and two indels were detected, and then five SNPs within different genomic regions were genotyped in the ISU Berkshire × Yorkshire pig resource family. The linkage disequilibrium analyses revealed that most of these FTO variants were not in strong LD with each other. The SNPs c.46–139A > T within intron 1 and a synonymous mutation c.594C > G (Ala198Ala) within exon 3 had significant (P < 0.01) associations with average daily gain on test and total lipid percentage in muscle, respectively. Five major haplotypes were identified and the subsequent association analyses suggested that haplotype 2 (-CTTGG-) was the most favorable for increased growth rate, while haplotype 1 (-CTACG-) was unfavorably associated with intramuscular fatness traits.     

Molecular Cloning and Expression Analysis of CD82 in Pig

CD82, which was originally referred to as KAI1 (kangai 1), is a member of the tetraspanin protein family, which contains four transmembrane domains. CD82 is implicated in a variety of biological processes, including apoptosis, cell adhesion, and cell migration. In this study, the full-length cDNA of pig CD82 was cloned and sequenced. Pig Cd82 cDNA contains an open reading frame (801 bp) encoding 266 amino acids. Sequence alignment results indicated that pig CD82 cDNA evidenced 85.45%, 85.63%, 77.03%, and 77.78% identity with human, cattle, rat, and mouse, respectively. In the expression study, the constitutive expression of swine Cd82 mRNA was detected in a variety of tissues, including lymphoid tissues as well as nonlymphoid tissues. Future studies will be focused on the functional role of CD82 during the course of pig infectious diseases or tumor development.