Carbonic Anhydrase Polymorphism in Cattle and Swine

Two electrophoretically different carbonic anhydrase (Ca) isoenzymes have been demonstrated in cattle where their presence is presumed to be controlled by codominant allelic genes (Sartore & Bernoco 1966). Previous reports of Ca polymorphism in swine are not available.     

奶牛和肉牛6个STR基因座遗传多态性研究

利用PCR技术和复合电泳银染技术检测奶牛和肉牛BM2113、BM1862、BMc701、BM2934、TGLA122、BM720等6个STR基因座的多态性分布,并计算该6个基因座的基因频率(P_i)、个体鉴别力(DP)、杂合度(H)、多态信息含量(PIC)、和非父排除概率(PE)。结果显示:6个STR基因座的基因型分布符合Hardy-Weinberg平衡,奶牛中6个STR基因座中BM2113基因座的DP、H和PIC最高,TGLA122基因座的PE最高。6个STR基因座的累积个体鉴别力(CDP)为0.99997,累积非父排除能力(CPE)为0.98827。肉牛中6个STR基因座中BM1862基因座的DP、H、PIC、PE都是最高,6个STR基因座的累积个体鉴别力(CDP)为0.99999,累积非父排除能力(CPE)为0.99578。结果表明,6个STR基因座可用于牛的遗传连锁分析、个体识别和亲子鉴定等研究领域。     

A High Throughput Single Nucleotide Polymorphism Multiplex Assay for Parentage Assignment in New Zealand Sheep

Accurate pedigree information is critical to animal breeding systems to ensure the highest rate of genetic gain and management of inbreeding. The abundance of available genomic data, together with development of high throughput genotyping platforms, means that single nucleotide polymorphisms (SNPs) are now the DNA marker of choice for genomic selection studies. Furthermore the superior qualities of SNPs compared to microsatellite markers allows for standardization between laboratories; a property that is crucial for developing an international set of markers for traceability studies. The objective of this study was to develop a high throughput SNP assay for use in the New Zealand sheep industry that gives accurate pedigree assignment and will allow a reduction in breeder input over lambing. This required two phases of development- firstly, a method of extracting quality DNA from ear-punch tissue performed in a high throughput cost efficient manner and secondly a SNP assay that has the ability to assign paternity to progeny resulting from mob mating. A likelihood based approach to infer paternity was used where sires with the highest LOD score (log of the ratio of the likelihood given parentage to likelihood given non-parentage) are assigned. An 84 “parentage SNP panel” was developed that assigned, on average, 99% of progeny to a sire in a problem where there were 3,000 progeny from 120 mob mated sires that included numerous half sib sires. In only 6% of those cases was there another sire with at least a 0.02 probability of paternity. Furthermore dam information (either recorded, or by genotyping possible dams) was absent, highlighting the SNP test’s suitability for paternity testing. Utilization of this parentage SNP assay will allow implementation of progeny testing into large commercial farms where the improved accuracy of sire assignment and genetic evaluations will increase genetic gain in the sheep industry.     

10个牛品种线粒体12S rRNA基因多态性分析

以鲁西牛、渤海黑牛、大别山牛、南阳牛、晋南牛、秦川牛、中国西门塔尔牛、日本和牛、海福特牛和安格斯牛10个牛品种共计353头牛为研究对象,利用单链构象多态性（SSCP）分子标记方法对其线粒体DNA 12S rRNA基因进行了多态性分析,结果发现该基因出现分布频率不同的3种单倍型（Ⅰ、Ⅱ、Ⅲ）,其中单倍型Ⅰ出现的频率最低,仅在晋南牛1个个体中发现;单倍型Ⅱ在中国本地黄牛中都存在,在中国西门塔尔牛中的分布频率很低,但在3个国外品种海福特牛、安格斯牛和日本和牛中不存在;单倍型Ⅲ在这10个牛品种中均有分布,但以国外品种最丰富。计算平均多态信息含量,发现我国地方品种在0.232～0.423之间,基本属于中度多态,说明我国地方黄牛品种在12S rRNA上多态性比较丰富,国外品种较为贫乏;但是单倍型Ⅰ频率很低有丢失的趋势,需要对其进行保护。     

奶牛TLR4基因的多态性与乳房炎的相关性研究

TLR4通过识别病原体而激活免疫细胞,在先天免疫和适应性免疫防御中起着重要作用。以中国荷斯坦奶牛、三河牛和中国西门塔尔牛为研究对象,以TLR4为乳房炎抗性的候选基因,分别扩增316bp和382bp2个片段,分别采用SSCP和RFLP-AluⅠ方法来检测TLR4基因的多态性。结合测序发现:在intron1的第4,525bp处的A→G的突变,和exon3的第1,397bp处的T→C突变,使得产生多态。2个位点的A、B等位基因在3个群体中都有分布,且处于中度多态。χ2适合性检验表明,3个群体在这2个位点的突变达到Hardy-Weinberg平衡状态(P>0.05)。运用SAS8.0软件采用最小二乘法拟合线性模型,将基因座不同基因型与奶牛乳房炎进行了关联分析,结果表明:T4CRBR1的AA基因型为乳房炎抗性基因型(P<0.05),A等位基因为乳房炎抗性的有利基因,T4CRBR2的各基因型个体间的体细胞评分差异不显著(P>0.05)。     

Purification of Rat Ceruloplasmin: Characterization and Comparison with Human Ceruloplasmin

Rat ceruloplasmin was purified by a three-step column chromatography procedure, utilizing DEAE-Sepharose, Sepharose CL-6B, and CM-Sephadex A50 columns. The molecular weight of rat ceruloplasmin determined by a molecular sieve column was 124,000 daltons. An optical density ratio (610 nm/280 nm) of 0.051 and a molar extinction coefficient of 8600 were obtained. A decrease in lysine in rat ceruloplasmin compared with human ceruloplasmin could account for its reduced anodal mobility. Other differences in the amino acid sequence of the rat ceruloplasmin included an increase in methionine and cystine/cysteine, and a decrease in histidine, tyrosine and tryptophan.     

Studies of Transferrin Polymorphism in Swedish Cattle Using Agarose Gel Electrophoresis

The polymorphic transferrin picture in the sera from 894 Swedish cattle was investigated with an agarose gel electrophoresis technique. The serum transferrin bands in the electrophoresis pattern were first identified by labelling with 59Fe. Six existing phenotypes based on the alleles TfA, TfD and TfE could be detected. The frequencies of transferrin types and transferrin alleles are presented, and it is concluded that there are great differences in the frequencies between the Swedish Red and White and the Swedish Friesian.     

甘肃西门塔尔牛甘州类群MSTN基因外显子多态性分析

采用PCR-SSCP方法检测50头甘肃西门塔尔牛甘州类群MSTN基因3个外显子的遗传多态性,并对群体内各等位基因进行测序,旨为探讨西门塔尔牛MSTN基因与肉质性状关联等方面的研究提供理论依据。结果显示,甘肃西门塔尔牛甘州类群MSTN基因3个外显子中,外显子2受B、C等位基因控制,形成BB、CC和BC 3种基因型,基因型频率分别为0.22(11/50)、0.64(32/50)、0.14(7/50);外显子1和外显子3分别受A等位基因与D等位基因控制,形成AA和DD基因型,基因型频率均为1(50/50)。序列分析表明,甘肃西门塔尔牛甘州类群MSTN基因3个外显子中,外显子2在第41 bp处发生了C→T的突变,但并没有导致氨基酸发生改变,属于同义突变;外显子1和3均无突变。统计结果表明,甘肃西门塔尔牛甘州类群MSTN基因3个外显子中,外显子2为中度多态,其余均无变化。     

Toll-Like Receptor 2 Gene Polymorphism and its Relationship with SCS in Dairy Cattle

Toll-like receptor 2 (TLR2) plays an important role in the innate immune response to a variety of pathogens. In this study, bovine TLR2 gene was taken as a candidate gene for mastitis resistance. Through PCR-SSCP analysis and sequencing, three missense mutations at T385 G, G398A, and G1884A were detected in the coding region that encoded extracellular domain. Altogether 240 dairy cattle of three breeds (Holstein, Simmental, and Sanhe cattle) were genotyped and allele frequencies were determined. The effects of TLR2 polymorphisms on somatic cell score (SCS) were analyzed and significant association was found between T385 G and SCS. The mean of genotype GG was significantly lower than those of genotype TT and TG. No significant associations were found with SCS for G398A and G1884A. Information provided in this research will be useful in further studies to determine the role of TLR2 gene in the mastitis resistance.     

奶牛MHC基因多态性及其与经济性状关系的研究进展

本文简述了奶牛 MHC基因的结构、位置、分类、多态性的一般特性,并且介绍了 MHC基因与生产性状、乳房炎等一些经济性状的关系。     

Ceruloplasmin and cardiovascular disease

Transition metal ion–mediated oxidation is a commonly used model system for studies of the chemical, structural, and functional modifications of low-density lipoprotein (LDL). The physiological relevance of studies using free metal ions is unclear and has led to an exploration of free metal ion-independent mechanisms of oxidation. We and others have investigated the role of human ceruloplasmin (Cp) in oxidative processes because it the principal copper-containing protein in serum. There is an abundance of epidemiological data that suggests that serum Cp may be an important risk factor predicting myocardial infarction and cardiovascular disease. Biochemical studies have shown that Cp is a potent catalyst of LDL oxidation in vitro. The pro-oxidant activity of Cp requires an intact structure, and a single copper atom at the surface of the protein, near His⁴²⁶, is required for LDL oxidation. Under conditions where inhibitory protein (such as albumin) is present, LDL oxidation by Cp is optimal in the presence of superoxide, which reduces the surface copper atom of Cp. Cultured vascular endothelial and smooth muscle cells also oxidize LDL in the presence of Cp. Superoxide release by these cells is a critical factor regulating the rate of oxidation. Cultured monocytic cells, when activated by zymosan, can oxidize LDL, but these cells are unique in their secretion of Cp. Inhibitor studies using Cp-specific antibodies and antisense oligonucleotides show that Cp is a major contributor to LDL oxidation by these cells. The role of Cp in lipoprotein oxidation and atherosclerotic lesion progression in vivo has not been directly assessed and is an important area for future studies.     

Polymorphism of intron 2 of the SDF1 gene in Galloway, Hereford, and Russian Black Pied cattle

The region of intron 2 of the SDF1 gene encoding a chemokine of the CXC subfamily has been resequenced in Galloway, Hereford, and Black Pied cattle. Five of the single-nucleotide polymorphisms (SNPs) that were earlier detected by other authors in various breeds of cattle in North America (99C/G, 128T/C, 206C/T, 267C/G and 313C/T) have been found. The 270insC polymorphic marker has proved to be monomorphic in Russian cattle breeds. Hereford cattle significantly differ from Galloway and Black Pied cattle in the frequencies of some SNP variants and their combinations. The number of SNP combinations in Hereford and Galloway cattle exceeds that in Black Pied cattle.     

Resistance of Brahman and Hereford Cattle to African ticks with reference to serum gamma globulin levels and blood composition

Field collections of ticks from two breeds of cattle showed that the common species of ticks wereAmblyomma hebraeum, Rhipicephalus appendiculatus andRhipicephalus evertsi evertsi. The density of these species was higher on Hereford than on Brahman cattle. The results also indicated that the density of immature stages of the above-mentioned ticks is higher on Hereford than on Brahman cattle.A positive correlation was found between the number of ticks on the cattle and the serum gamma globulin levels, an indication of an increase in the production of antibodies. An inverse relationship was found between tick burden and red blood cell count and hemoglobin concentration. Other ablood components, such as basophils, eosinophils and lymphocytes were not affected by the changes in the density of the tick populations. It appears that resistance may have been acquired by the hosts and that Brahman cattle may acquire resistance to a higher degree than Herefords.     

Ceruloplasmin immunoreactivity in neurodegenerative disorders

Ceruloplasmin (CP) is a 132kd cuproprotein which, together with transferrin, provides the majority of anti-oxidant capacity in serum. Increased iron deposition and lipid peroxidation in the basal ganglia of subjects with hereditary CP deficiency suggest that CP may serve as an anti-oxidant in the brain as well. The present study compared CP immunoreactivity in brain specimens from normal controls and subjects with neurodegenerative disorders (Alzheimer's disease [AD], Parkinson's disease [PD], progressive supranuclear palsy [PSP], and Huntington's disease [HD]) (n = 5 per group). The relative intensity of neuronal CP staining and the numbers of CP-stained neurons per 25x microscope field were determined in hippocampus (CA1, subiculum, and parahippocampal gyrus), parietal cortex, frontal cortex, substantia nigra, and caudate. CP was detected in both neurons and astrocytes in all specimens, and in senile plaques and occasional neurofibrillary tangles in AD brain. Neuronal CP staining intensity tended to increase in most AD brain regions, but was statistically significant vs controls only in the CA1 region of hippocampus (p = .016). Neuronal CP staining in brain specimens from other neurodegenerative disorders showed a slight but nonsignificant increase vs controls. The numbers of CP-stained neurons per field did not differ between the various neurodegenerative disorders and controls. These results suggest that a modest increase in neuronal CP content is present in the AD brain, and lesser elevations in neuronal CP occur in the other neurodegenerative disorders in this study. Though CP functions as both an acute phase protein and an anti-oxidant in peripheral tissues, whether it does so in the brain remains to be determined.     

铜蓝蛋白与脑铁代谢

铜蓝蛋白（Cerulopasmin,CP)是人体重要的亚铁氧化酶。它的主要作用是催化二阶铁成为三阶铁,从而促进铁与转铁蛋白结合。由于它的