香猪基因组外显子SNPs检测及繁殖和体型性状候选基因分析

为从基因组水平上探究香猪性成熟早、产仔数少、体型小的分子机理,本研究下载与香猪在繁殖和体型性状方面有明显差异的梅山猪、杜洛克猪的重测序数据作为参照,对26头香猪、24头梅山猪和24头杜洛克猪的重测序数据进行SNPs检测和等位基因频率差值分析,筛选香猪基因组外显子上与梅山猪、杜洛克猪高度分化的SNPs位点,并使用AS-PCR方法和Sanger测序方法统计其中8个SNPs位点的群体等位基因频率。最后,对含有高度分化SNPs位点的基因进行GO功能富集和KEGG通路分析。结果显示,SNPs检测获得香猪基因组外显子上SNPs共236 710个,包括193个终止密码子丢失、1 238个终止密码子获得、90 945个错义SNPs和144 334个同义SNPs。得到1 046个香猪高度分化SNPs,包括429个错义SNPs、2个终止子丢失、6个终止子获得和609个同义SNPs,影响524个蛋白质编码基因;并发现X染色体上44-58 Mb的一段富含高度分化SNPs的热点区域。群体等位基因频率验证结果与重测序结果一致,鉴定了8个位点均为香猪与梅山猪、杜洛克猪高度分化的SNPs。对524个含有高度分化SNPs的基因进行GO和KEGG分析,富集到卵母细胞减数分裂、雌激素信号途径等繁殖相关通路;GO功能注释到细胞内雌激素受体信号、纤毛运动等繁殖相关条目,骨骼系统形态发生、骨骼发育、成骨细胞分化等体型相关条目。得到含有香猪高度分化SNPs的18个繁殖性状候选基因PTGFR、TRO、DNAH17、PKDREJ、KAT8、DNAI2、VDR、TEX14、QSOX1、AR、WNK3、ADCY3、SPEF1、MIGA2、SLC2A8、PSMF1、TBC1D20、IFT172和7个体型相关基因IBSP、NR6A1、HSPG2、TARS、PDZD2、FGF4、AMER1,可能是决定香猪性成熟早、产仔数少、体型小的关键基因。     

Cloning of Inhibin-α Gene and its mRNA Expression Pattern in the Ovary of Congjiang Xiang Pig

To reveal the relationship between inhibin-α(INHA) gene and the reproductive traits of Congjiang Xiang pig, INHA gene was cloned and sequenced taking the genomic DNA of Congjiang Xiang pigs as templates by polymerase chain reaction(PCR) method.The polymorphisms of INHA gene were tested in Congjiang Xiang pig populations with high-litter size and low-litter size using allele-specific PCR(AS-PCR) method.The expression profile of INHA gene in ovaries was detected from Congjiang Xiang pigs with high-litter yiled or low-litter yiled by Real-time PCR method.The complete coding region of INHA gene was 1095 bp in length, which coded for 364 amino acid residues.Compared with the known sequence, two candidate sites, G359A and A373G, were found out from exon 2 region of INHA gene in Congjiang Xiang pig.After investigation for the two sites in a large population, the frequency of alleles between two populations was not significant and without obvious relativity with the litter yiled of Congjiang Xiang pig.However, the INHA mRNA level in the ovary of Congjiang Xiang pig with high-litter yiled was higher than that with low-litter yiled.It suggested that INHA gene was much conserved, INHA gene expression level might be concerned for the regulation of ovary growth and follicle development in Congjiang Xiang pig breed.     

Isolation and Identification of BPIV3 Genotype C

To identify the infection agents from Ningxia Hui Autonomous region, where feedlot cattle indicated bovine respiratory disease complex (BRDC), the M gene of the bovine parainfluenza virus type 3 was amplified by RT-PCR.The PCR product was ligated to pMD18-T vector and cloned to E.coli DH5α.The positive clones were sequenced and compared with the reference strains in GenBank by the molecular biology software.Sequence alignment results showed that a BPIV3 strain was isolated from the samples and named NX49, the M gene of NX49 included 1 056 nucleotides.Evolutionary analysis showed that the NX49 belonged to BPIV3 C genotype and shared 99.4% nucleotide identity with that of the SD0835 isolated in Shandong province.The characterization of the NX49 demonstrated that it was sensitive to temperature, acid and organic matter.The presence of Mg²⁺ showed no protection against the treatment at high temperature.The HA test suggested that the NX49 enables to agglutinate the guinea pig RBC at 4 ℃ and the titer was 1∶4.The study isolated a BPIV3 genotype C strain successfully, which facilitate the study of molecular evolution and epidemiology of BPIV3 in China.     

Analysis on Polymorphisms of Three CNV Regions in Five Pig Breeds

The paper was aimed to investigate the polymorphism of copy number variation (CNV) in different pig breeds.Three CNV regions of CNVR91,CNVR92 and CNVR143 were chosen from the porcine SNP60 chip genotyping results.The polymorphisms of three CNVs were determined by Real-time quantitative PCR method,taking five pig breeds as samples,including Yorkshire pig,Xiang pig,Kele pig,Nuogu pig and Rongchang pig breeds.The results showed that the dominant status of CNVR91 was loss in Xiang pig,while it was normal in other four pig breeds.The major type of CNVR92 was deletion in Xiang pig,Yorkshire pig,Kele pig and Rongchang pig breeds,with a high normal percent in Nuogu pig.For CNVR143,the dominant event was gain in Xiang pig and Nuogu pig breeds,but it was not diverse in other three pig breeds.These results indicated that three CNV regions emerged with polymorphism in five pig breeds,which might have effects on gene expression in CNV regions and physiological function by dosage effect especially in Xiang pig,Nuogu pig and Kele pig breeds.     

Inhibition of porcine circovirus type 1 and type 2 production in PK-15 cells by small interfering RNAs targeting the Rep gene

Porcine circovirus type 1 (PCV1) and type 2 (PCV2) are two genotypes of porcine circovirus. Both of them are presumed to be widespread in the swine population. Currently, there is no specific treatment for their infections. RNA interference (RNAi) is a sequence-specific RNA degradation mechanism mediated by small interfering RNA (siRNA), which represents a possible therapeutic application for the treatment of viral infections. In this study, three siRNA expression plasmids (pS-RepA, pS-RepB and pS-RepC) were generated to target three different coding regions of the Rep protein (Rep) of PCV. These siRNAs were used to inhibit PCV production in a porcine kidney cell line, PK-15 cells. Our results revealed that Rep gene expression was inhibited by pS-RepA, pS-RepB and pS-RepC to different degrees. Moreover, our study also showed that the production of PCV1 and PCV2 was reduced by these siRNAs. pS-RepC, which targets the middle region of Rep gene, proved to be the most efficient siRNA for inhibition of Rep expression and viral production. Taken together, our data suggest that RNAi could be investigated as a potential treatment for PCV infection.     

管理措施对三江源区"黑土滩"土壤肥力及土壤酶活性的影响

对三江源区典型退化草地“黑土滩”采取施肥及种植垂穗披碱草的恢复措施,结果表明,土壤中的全氮与有机质的相关性显著,全磷与有机质的相关性较明显,变化趋势相一致。短期的恢复措施造成了恢复样地20~30 cm土壤层中有机质、全氮和全磷含量的下降,长期的恢复措施使恢复样地中的这3种营养物质的含量得到了上升;人工种植措施使土壤中的全钾含量显著下降(P<0.05)。此外,速效养分的变化不一致,其中速效氮的含量表现为先降低后升高,速效磷和速效钾的含量变化表现出升高趋势。植被恢复对土壤脲酶活性有促进作用,但蔗糖酶活性没表现出规律性变化;这2种酶的活性与速效养分的含量表现出较强的正相关,而与有机质和全价养分的含量相关性不显著。     

Reuolution and Research of Green Automobile

The green is as a symbol of no pollution,no public harm and environmental protection. The green automobile is the inevitable results that the automobile technique continuously developes. A new style vehicle--the green automobile is the urgent solicit of environmental protection is developed. The automobile industry can develop continuously. This article mainly discusses the importance of the green auto_mobile's research and development,the development status in quo and prospect of the green auto_mobile technique in the world,and states the relationship between the green automobile technique and environmental protection from green technical system theoretically clarify the study of green automobile,including its design and the system,the manufacture and its technology,the energy,the rebuild engineering and a series of subjects. How to evaluate the cycle and the evaluable mark of the green automobile,the prospect of green automobile are pointed out.     

An Estimate on the Casual Relation between Financial Development and Economic Growth

This paper firstly discusses the relationship among money,real interest rate and economic growth of China. Then estimates the causality between financial development and economic growth by Bivariate Granger Casual Relation Test. The conclusion suggests that economic growth stimulate financial development more in China.     

A New Adaptive Sequential Diagnosis Algorithm in the PMC Model

We present an adaptive algorithm for sequential diagnosis in the PMC model,which can directly identify a faulty processor without first identifying a fault-free processor.     

The Inherent Fault Tolerance of Multi-Layer Neural Networks

The inherent fault tolerance of multilayer neural networks (MLNs) is due to thedistributed properties of information processing and information storage in an MLN , and the inher-ent fault-masking capability of every neuron. Also,the concept of inherent fault tolerance of MLNsis formalized, Finally, perceptrons and three-layer nets are shown inherently fault tolerant underreasonable statistical conditions,respectively.