Study on Developmental Expression of ApoCⅡ Gene mRNA in Liver Tissues of Pigs

The aim of this study was to investigate the developmental patterns of ApoCⅡ gene mRNA in liver in Mashen and Large White pigs, and study the relationship between the expression level of ApoCⅡ and the lipid metabolism in pigs.The mRNA relative expressions of ApoCⅡ gene in liver at seven stages of 1,30,60,90,120,150,and 180-day old in Mashen and Large White pigs were determined by quantitative Real-time PCR.The results showed that the developmental trend of ApoCⅡ mRNA expression in liver between Mashen and Large White pigs was different.The ApoCⅡ mRNA abundance was decreased from birth to 60-day old,then increased at 90-day old,and decreased again after that in Mashen pig.However,the relative expression amount in Large White pig was gradually decreased from birth to 150-day old and increased again at 180-day old.Except for the ApoCⅡ mRNA expression amount at 1-day old,the differences of the expression amount at other stages in Mashen and Large White pigs were significant or extremely significant (P<0.05 or P<0.01).The ApoCⅡ mRNA expression in liver was affected by age and breed,and could play an important role in lipid metabolism in pigs.     

重组猪抑制素蛋白质中毒素残留检测及其理化稳定性

为探索纯化的重组猪抑制素蛋白质毒素残留情况以及重组猪抑制素蛋白质稳定性,本研究利用鲎试剂、气相色谱和SDS-PAGE方法对重组猪抑制素蛋白质中的细菌内毒素、β-巯基乙醇的残留量及其蛋白质在不同温度、极端p H值和反复冻融条件下的稳定性进行检测。结果显示,经过3次等电点沉淀洗涤后,重组猪抑制素蛋白质中细菌内毒素和β-巯基乙醇的残留量大幅减少。通过在不同温度下的处理发现,重组猪抑制素蛋白质在4℃环境下较稳定,可以长期保存;但在37℃时,稳定性快速下降,随着温度上升而降解加速。通过极端p H值和反复冻融试验发现,重组猪抑制素蛋白质对极端p H值和反复冻融具有较好的耐受性。     

Amplification,Sequence Analysis and Tissue Expression Detection of FUT2 Gene Coding Regions of Dongchuan Pigs

Porcine alpha (1,2) fucosyltransferase (FUT2) gene was importance in glycosphingolipid biosynthesis-globo series, potentially played a regulatory role during Escherichia coli (E. coli) F18 infection process in weaned piglets. In order to explore sequence structure of porcine FUT2 gene and its biological function, this test amplified FUT2 gene CDS sequence of Dongchuan pigs by PCR, forecasted and analyzed the protein sequences and functional regions of FUT2 gene, its expression level was detected in 11 tissues of 8 Dongchuan weaned piglets in 35 days old at the meantime. The results showed that the CDS sequence of FUT2 gene was 1 023 bp, which encoded 340 amino acids. FUT2 protein was fat-soluble hydrophilic protein, which the structure was not stable, including a transmembrane helix structure, but without signal peptide that suggested the FUT2 protein was a membrane protein;FUT2 protein included 2 N-glycosylation sites (No. 185 and No. 305 amino acids), without O-glycosylation sites, there were 14 potential phosphorylation sites, included 6 Ser, 2 Thr and 6 Tyr, analyzing the functional regions found that the FUT2 protein had a superfamily of conserved domains:FUT1-FUT2-like (58-319 amino acids). The phylogenetic tree result showed that the relative relationship between swine and cattle was relatively close, but was distant from chimpanzee, human, mouse and rat. FUT2 gene was expressed in all 11 tissues of Dongchuan weaned piglets, there were higher expression in digestive tract and immune tissues. The present results suggested that FUT2 gene might play a role to resistance to E. coli F18 in weaned piglets, and might indirectly against E. coli F18 through the synthesis of fucosyltransferase.     

Cloning and Sequence Analysis of ACOX1 Gene in Luchuan Pig

The objective of this study was to clone the acyl-coenzyme A oxidase 1 (ACOX1) gene of Luchuan pig and analyze its genetic structure with bioinformatics. A pair of special primer was designed according to predicted sequence of pig ACOX1 gene in GenBank. The ACOX1 gene was amplified by RT-PCR, the physicochemical property and secondary structure of ACOX1 gene were systemically analyzed by bioinformatics techniques. The results showed that ACOX1 gene fragment included an 1 986 bp whole length CDS (coding 661 bp amino acids). The sequence multi-aligned results showed that Luchuan pig shared 99.5%, 85.5%,87.1%,66.3%,75.6%,84.0%,82.3%,81.1% and 69.1% of similar nucleotide sequence with that of pig,human, zebrafish, chicken, rhesus monkeys, mice, rat and frog, respectively. The prediction of ACOX1 secondary structure showed that Luchuan pig ACOX1 had no signal peptide and transmembrane structure. This study suggested that the whole CDS sequence of ACOX1 gene was successfully cloned in Luchuan pig, and the cloning and analysis of ACOX1 gene provided an important foundation for further studying on the fatty deposition and lipornetabolism of ACOX1 gene in Luchuan pig.     

Characterization of MC1R and KIT Genotypes in Luchuan Pig Population

In order to assess the purity of Luchuan pig populations, four South Chinese local pig breeds including Putian Black pig, Yuedong Black pig, Dahuabai pig, Bama miniature pig and three foreign pig breeds including Duroc pig, Yorkshire pig and Landrace pig were studied as controls by sequencing of PCR products, MC1R and KIT genotypes in 56 Luchuan pigs were analyzed in this study. Sequencing results indicated that a splicing mutation (G>A) was presented in the first base in intron 17 of KIT gene in both Yorkshire pig and Landrace pig, in contrast, the wildtype GG of KIT gene was presented in Luchuan pig, four south Chinese local pig breeds and Duroc pig.Compared with Hainan wild boar, South Chinese local pig breeds had two missense mutations 95Val > Met and 102Leu > Pro in the coding region of MC1R gene;Compared with Yorkshire pig, Landrace pig and Duroc pig, South Chinese local pig breeds had 5 to 6 SNPs in MC1R gene 5'UTR, and in addtion, an A base deletion in MC1R gene 3'UTR. Furthermore, we found one litter of Luchuan pig with abnormal coat color.The results showed that the presentation of two distinct MC1R genotypes E^D1 and E^p in both litters and the sow,but only E^D1 in the boar. Considering E^p was derived from Pietrain pig, we preliminarily considered that the genome of the sow might be infiltrated with foreign pig breeds. In summary, we detected the genotypes of the coat color genes KIT and MC1R in eight pig breeds, confirmed the molecular differences of coat color between Chinese local pig breeds and foreign pig breeds, which could be useful for the further investigation of the molecular mechanism of pig coat color.     

Differential Expression Analysis of MUC4 and MUC13 Genes Between Resistant and Sensitive Weaned Meishan Piglets to ETEC F18

MUC4 and MUC13 genes as important candidate genes for enterotoxigenic Escherichia coil (ETEC) F4 resistance,may play an important role in the process of against ETEC F18 infection in weaned piglets. In this study,ETEC F18-resistant and -sensitive weaned Meishan piglets were used,and the expression levels of MUC4 and MUC13 genes in 11 tissues (heart,liver,spleen,lung,kidney,stomach,muscle,thymus,lymph nodes,duodenum and jejunum) were determined by quantitative Real-time PCR. The results showed that MUC4 and MUC13 genes were broadly expressed with different expression levels in all the 11 tissues. In the thymus and lymph tissues,the expression of MUC4 gene in resistant piglets was significantly higher than that in sensitive piglets (P<0.05);In the lung tissue,theMUC13 gene expression level in resistant individuals was significantly higher than that in sensitive individuals (P<0.05),and in the intestinal tissues of duodenum and jejunum, the expression level of MUC13 gene was relatively higher in resistant individuals. Thus we speculated that the high expression of MUC4 gene in immune tissues and MUC13 gene in intestinal tissues might improve the immune ability of piglets,protect and lubricate the intestinal tract, and resist ETEC F18 infection.     

Research Progress and Countermeasures of Porcine Endogenous Retrovirus in Chinese Miniature

The paper is a brief introduction of porcine endogenous retrovirus (PERV) innovative research results in the world and also expounds the passing PERV existence situation on different varieties of miniature pig, analyzes the PERV-virus gene cloning and sequence, appraises method on cell level, create the platform of infection HEK293 cells research, study on pigs A3F inhibition of PERV, and reveal the innovative research results on the specific molecular genetics in the different strain of PERV, analyzes the advantages of Wuzhishan miniature pig inbred line such as low gene copy of PERV,and there is no passing PERV-C specificity and as well as looking forward to cultivate the methods for new strain of PERV negative pigs. It will provide a scientific counter measure and new perspective to solve the spread of disease risk of miniature pig PERV and product safety for human xenotransplantation and biomedical materials of research and development.     

Cloning and Sequence Analysis of PTTG1 Gene in Luchuan Pig

This experiment was aimed to clone PTTG1 gene CDS sequence of Luchuan pig,and was analyzed by bioinformatics methods.A pair of special primers was designed according to predicted sequence of porcine PTTG1 in GenBank.The coding sequence of PTTG1 in Luchuan pig was amplified by RT-PCR,its gene sequence characteristics and protein structure was systemically analyzed by bioinformatics techniques.The results showed that the cloned PTTG1 fragment included a 609 bp CDS (coding 202 amino acids).The sequence multi-aligned results showed that Luchuan pig shared 90.15%,87.85%,87.52%,87.03%,76.03%,74.38%,55.74% and 44.48% of similar nucleotide sequence with that of Bos,Pan troglodytes,Homos,Macaca,Rattus,Mus,Gallus and Danio retio,respectively.The protein structure analysis results showed that the protein attributed hydrophilic protein without signal peptide,localized in cell cytoplasm and had 16 phosphorylation sites.The phylogentic tree of amino acid indicated that PTTG1 was highly conserved in the process of evolution of different species.The cloning and analysis of PTTG1 gene provided an important foundation for further study biological function of porcine PTTG1 during early embryonic development.     

山东省生猪生产格局及其影响因素的实证分析

依据《山东统计年鉴》(1981—2012)中的相关统计数据,阐述了20世纪80年代以来山东省生猪生产格局变化的历史进程;在理论分析生猪生产格局影响因素的基础上,应用面板数据模型实证分析山东省生猪生产格局的影响因素。结果表明:1982年以来山东省生猪生产格局经历了从"√"型格局到"⌒"状格局再到"·—S"格局的变化过程;上一年生猪生产格局指数、消费者市场潜力、政策扶持和畜牧业比较优势对生猪生产格局变化有显著正影响,非农就业机会有显著负影响,且上一年生猪生产格局指数的影响程度明显高于其余4个因素。     

系统分析多组织转录组鉴定影响猪脂肪沉积的关键基因

旨在挖掘影响松辽黑猪脂肪沉积的关键基因及脂肪、肝和肌肉在体内的功能。本研究选择体重100 kg左右健康且背膘厚差异显著的6头(高、低各3头)松辽黑猪为试验动物,利用高通量转录组测序技术检测其脂肪、肝和背最长肌组织中基因的表达水平,鉴定不同脂肪沉积猪和不同组织中的差异表达基因,并分析差异表达基因的生物学功能。结果表明,在不同分组的猪中发现135个差异表达基因,其中部分参与了PPAR信号通路、AMPK信号通路、代谢通路、脂肪酸代谢和甘油代谢等通路。经生物学功能分析发现,EHHADH、ME1、SCD、OLR1、PHGDH、ACLY、LEP和CYP超家族基因等基因为影响猪脂肪沉积的关键基因。在不同组织的差异表达表达基因中,脂肪组织中高表达的基因显著富集在胰岛素信号通路、MAPK信号通路、三羧酸循环、氧化磷酸化等通路;肝中高表达基因显著富集在多种物质的代谢、脂肪酸的降解、氨基酸的合成等通路;背最长肌中高表达的基因主要参与了蛋白质的降解、PI3K-Akt信号通路、氧化磷酸化通路、Wnt信号通路、磷脂酰肌醇信号通路等通路。不同组间差异表达基因分析结果提示,EHHADH、ME1、SCD、OLR1、PHGDH、ACLY、LEP和CYP超家族基因等基因是影响脂肪沉积的关键基因;不同组织间差异表达基因表明,脂肪组织是脂肪合成的主要部位,而肝和肌肉组织主要涉及脂肪酸的降解。本研究结果对脂肪性状的遗传改良、机理解析有一定意义。