Genetic variation in two conserved local Romanian pig breeds using type 1 DNA markers

Analysis of the genetic variation of an endangered population is an important component for the success of conservation. Animals from two local Romanian pig breeds, the Mangalitsa and Bazna, were analysed for variation at a number of genetic loci using PCR-based DNA tests. Polymorphism was assessed at loci which 1) are known to cause phenotypic variation, 2) are potentially involved in trait differences or 3) are putative candidate genes. The traits considered are disease resistance, growth, coat colour, meat quality and prolificacy. Even though the populations are small and the markers are limited to specific genes, we found significant differences in five of the ten characterised loci. In some cases the observed allele frequencies were interesting in relation to gene function and the phenotype of the breed. These breeds are part of a conservation programme in Romania and marker information may be useful in preserving a representative gene pool in the populations. The use of polymorphisms in type 1 (gene) markers may be a useful complement to analysis based on anonymous markers.     

Detection of flavor compounds in longissimus muscle from four hybrid pig breeds of Sus scrofa,Bamei pig,and Large White

To detect the flavor quality and flavor compounds in raw longissimus muscle from four typical pig breeds: Sus scrofa?×?Bamei pig named F1 (group A), F1?×?F1 (group B), F1?×?Bamei pig (group C), and F1?×?Large White (group D). The chemical compositions of longissimus muscles from four breeds were examined using headspace solid-phase microextraction/gas chromatography mass spectrometry method. Distinct differences for the same flavor compounds of longissimus muscles between different breeds were analyzed. Totally 64 flavor compounds shared in four groups, and 10 flavor compounds with significant difference among four groups (p?<?0.05), including allyl butyrate, (Z)-2-penten-1-ol, 2,2-dimethyl-3-methyl oxirane, 2-pentylfuran, dodecane, 2,4-decadienal, vinylsilane, 3-methyl-1-butanol, (1-methyldecyl)-benzene, and dipropyl phthalate. Totally, 23–41 flavor compounds did not commonly exist in four groups, such as only as dibutyl isophthalate in group A; 6,10-dimethyl-5-9-undecadien-2 one, bis (2-trimethylsilyl) ethyl ester-malonic acid, heptadecane, 2,4,6-trimethyl pyridine, and diisooctyl adipate in group C alone; and 1,3-dimethylcyclopentanol, 2-octanone, and trimethylsilane in group D alone. While, no specific flavor compounds were identified in group B. All these flavor compounds covered 12 types of hydrocarbons, alcohols, aldehydes, hydroxybenzenes, acids, ketones, esters, sulfides, furans, alkenes, and pyrrole. Besides, we analyzed 14 flavor compounds with different flavors combining with previous studies. The flavor compounds in longissimus muscles might be closely related to the breeds, and the hybrid of S. scrofa?×?Bamei pig had the most flavor compounds in raw longissimus muscle.     

Expression profiling analysis for genes related to meat quality and carcass traits during postnatal development of backfat in two pig breeds

The competitive equilibrium of fatty acid biosynthesis and oxidation in vivo determines porcine sub-cutaneous fat thickness(SFT) and intramuscular fat(IMF) content.Obese and lean-type pig breeds show obvious differences in adipose deposition;however, the molecular mechanism underlying this phenotypic variation remains unclear.We used pathway-focused oligo microarray studies to examine the expression changes of 140 genes associated with meat quality and carcass traits in backfat at five growth stages(1―5 months) of Landrace(a leaner, Western breed) and Taihu pigs(a fatty, indigenous, Chinese breed).Variance analysis(ANOVA) revealed that differences in the expression of 25 genes in Landrace pigs were significant(FDR adjusted permutation, P<0.05) among 5 growth stages.Gene class test(GCT) indicated that a gene-group was very significant between 2 pig breeds across 5 growth stages(PErmineJ<0.01), which consisted of 23 genes encoding enzymes and regulatory proteins associ-ated with lipid and steroid metabolism.These findings suggest that the distinct differences in fat deposition ability between Landrace and Taihu pigs may closely correlate with the expression changes of these genes.Clustering analysis revealed a very high level of significance(FDR adjusted, P<0.01) for 2 gene expression patterns in Landrace pigs and a high level of significance(FDR adjusted, P<0.05) for 2 gene expression patterns in Taihu pigs.Also, expression patterns of genes were more diversified in Taihu pigs than those in Landrace pigs, which suggests that the regulatory mechanism of micro-effect polygenes in adipocytes may be more complex in Taihu pigs than in Landrace pigs.Based on a dy-namic Bayesian network(DBN) model, gene regulatory networks(GRNs) were reconstructed from time-series data for each pig breed.These two GRNs initially revealed the distinct differences in physiological and biochemical aspects of adipose metabolism between the two pig breeds;from these results, some potential key genes could be identified.Quantitative, real-time RT-PCR(QRT-PCR) was used to verify the microarray data for five modulated genes, and a good correlation between the two measures of expression was observed for both 2 pig breeds at different growth stages(R=0.874±0.071).These results highlight some possible candidate genes for porcine fat characteristics and provide some data on which to base further study of the molecular basis of adipose metabolism.     

猪脂肪及肌肉组织中基因表达信息分析

为探测猪脂肪及肌肉组织中基因表达概况,利用猪EST资源和人类基因序列开展计算机模拟研究,旨在为猪肉质改良的遗传基础分析提供候选信息。执行Blast比对程序以识别人类基因组基因与猪EST序列间的同源性并筛选出高度同源记录,同时编制4个Java程序进行序列检索收集、序列比对结果的过滤筛选以及分类处理。统计分析表明：至少有2002个基因在猪脂肪及肌肉组织中表达,其中1087个基因在脂肪组织表达,1205个基因在肌肉组织中表达,两组织共同表达的基因为290个;筛选出高同源基因,同时分类统计出了114个基础活性基因(脂肪和肌肉组织分别表达80和34个),并选取Top记录进行了描述分析和总结。     

猪的骨骼肌生长发育研究进展

瘦肉率对生猪产业来说是一个极其重要的经济指标,而这一指标完全取决于骨骼肌的生长发育。因此,猪骨骼肌生长发育机理的研究是十分必要的。然而,在早期由于各种因素的限制,猪骨骼肌单个基因的研究一直进展缓慢;相反,以小鼠为模型,其骨骼肌单基因的功能研究却取得了较大进展。在这一时期,影响肌决定和肌分化的基因,如MRFs家族和MEF2家族相继被发现,这些基因在猪的肌肉发育中也发挥着同样的作用。然而,这些结果并不能很好地揭示骨骼肌发育过程中复杂的基因间互作关系。随着近年来芯片和测序技术的不断发展,更多人试图从整个转录谱的水平来阐述猪肌肉发育的分子机理,并且也取得了较大的进展。为了对猪骨骼肌生长发育有一个更为清晰的认识,该文将以目前猪骨骼肌生长发育研究结果为基础,同时结合模式动物小鼠骨骼肌单基因的研究成果,对猪的骨骼肌生长发育分子调控机理进行详细的阐述。     

The swine histo compatibility system SLA: serological studies in the common Swiss and Danish swine breeds

Alloantisera were produced in common Swiss and Danish swine breeds by immunization with leucocytes or skin-grafts. Out of the 126 antisera, 66 were chosen for further study based on titrations employing lymphocytes from unrelated pigs typed with French SLA antisera (Vaiman, Chardon & Renard, 1979). The 66 selected antisera and the French reagents defining 26 SLA specificities were used to type lymphocytes from 595 unrelated pigs of the common Swiss and Danish breeds. The reaction-patterns of the French, Swiss and Danish antisera were adequately correlated for the French SLA specificities Nos FJ 2, 3, 6, 7, 8, 9, 11, 14, 19, 20 and 24 and for the haplotypes FJ 15.1.18 and FJ 5.4. In addition, a cluster of correlated Danish and Swiss antisera characterized a new specificity, provisionally designated CPH 31. This specificity was frequent in the Danish Landrace pigs.
Using the reagents identified in this report, the segregation of SLA markers was studied. Back-cross families demonstrated segregation of 15 distinct SLA haplotypes of which 14 are common in French Landrace or Large White. Differences were found in haplotype frequencies in both the Swiss and the Danish Landrace and Large White breeds.     

不同猪品种肌肉组织FoxO1与MyoD基因mRNA的表达及其相关性分析

分别取6月龄八眉猪、长白猪和长×八杂交猪背最长肌,提取总RNA,设计并合成猪FoxO1和MyoD 引物,以β肌动蛋白作为内参,优化反应条件和体系,利用RT-PCR方法检测八眉、长白和长×八杂交猪肌肉组织中FoxO1和MyoD基因mRNA 的表达.结果表明,在不同经济类型猪品种肌肉组织中,FoxO1和MyoD基因mRNA的表达丰度均存在显著差异,FoxO1在八眉猪肌肉组织中的表达普遍高于长白猪（P<0.01）,在杂交组合长×八肌肉组织中的表达也高于长白猪（P<0.01）;而MyoD恰好相反,即在长白猪肌肉组织中的表达普遍高于八眉猪（P<0.01）,在杂交组合长×八肌肉组织中的表达也高于八眉猪（P<0.01）．结果提示,FoxO1和MyoD基因mRNA的表达在肌肉发育中存在负相关（r = 0.728 , P < 0.05）,FoxO1在肌肉组织中的上调作用,可能是造成的MyoD基因mRNA表达降低的原因之一,进而负调控肌肉的发育和骨骼肌的量．     

猪背最长肌中肌肉生长和脂肪沉积相关基因的差异表达和主成分分析

骨骼肌细胞和脂肪细胞在分化生长速度上相对竞争的平衡点是猪肉质和胴体性状的决定因素.利用Oligo功能分类芯片检测了瘦肉型的长白猪和脂肪型的太湖猪在初生、1、2、3、4和5月龄间背最长肌中肌肉生长和脂肪沉积相关基因的动态表达变化.差异表达分析结果显示,在初生至5月龄的品种间分别有15、16、11、13、18和20个基因的表达差异倍数大于2倍.品种内的方差分析表明,长白猪分别有18和22个基因,太湖猪分别有3和7个基因在月龄间的表达差异达极显著(Ｐ<0.01)和显著水平(Ｐ<0.05).主成分分析结果显示,先降后升是两品种内最具代表性的基因表达模式,且长白猪和太湖猪分别有7和6个基因的表达模式明显偏离其他基因,提示其可能受到了重要的调控. 此外,5个差异表达基因的荧光定量RT-PCR验证结果均与芯片结果呈正相关趋势.以上结果筛选出了对于猪肉质和胴体性状可能具有重要影响,值得深入研究的一些候选基因,为深入研究生长发育过程中参与肌纤维生长和脂肪酸合成关键基因的表达变化规律和互作调控机制提供了基础数据.     

Gene expression profiling of p53-sensitive and -resistant tumor cells using DNA microarray

Overexpression of wild-type p53 in ECV-304 tumor cells induced extensive apoptosis and the eventual death of nearly all of the cells. We generated ECV-304 cells resistant to p53-induced apoptosis as a strategy to identify novel genes that might be relevant to p53-mediated apoptosis. ECV-304 cells resistant to p53 were isolated by repeated infections with a recombinant p53 adenovirus and were designated as DECV. The expression of 5,730 genes in p53-resistant (DECV) and p53-sensitive ECV-304 cells were profiled by DNA microarray analysis. We report here the expression of 80 genes that differed by 2-fold or more between sensitive and resistant cells upregulated for p53. Many of these differentially expressed genes are regulated by p53 in ECV-304 and H1299 p53-null cells. Our analysis identifies many new potential targets for p53 that play roles in cell cycle regulation, DNA repair, redox control, cell adhesion, apoptosis, and differentiation.     

Protein profiling in human sera for identification of potential lung cancer biomarkers using antibody microarray

To identify potential biomarkers of lung cancer (LC), profiling of proteins in sera obtained from healthy and LC patients was determined using an antibody microarray. Based on our previous study on mRNA expression profiles between patients with LC and healthy persons, 19 proteins of interest were selected as targets for fabrication of an antibody microarray. Antibody to each protein and five nonspecific control antibodies were spotted onto a hydrogel‐coated glass slide and used for profiling of proteins in sera of LC patients in a two‐color fluorescence assay. Forty‐eight human sera samples were analyzed, and expression profiling of proteins were represented by the internally normalized ratio method. Six proteins were distinctly down‐regulated in sera of LC patients; this observation was validated by Wilcoxon test, false discovery rate, and Western blotting. Blind test of other 32 human sera using the antibody microarray followed by hierarchical clustering analysis revealed an approximate sensitivity of 88%, specificity of 80%, and an accuracy of 84%, respectively, in classifying the sera, which supports the potential of the six identified proteins as biomarkers for the prognosis of lung cancer.