猪繁殖与呼吸综合征病毒双基因多组合DNA疫苗的构建

本实验根据PRRSV结构蛋白的特性和IFNγ及IL-2这两种细胞因子的特性,构建表达PRRSVGP4-GP5、GP5-M、GP5-N、GP5-IFNγ、GP5-IL-2、N-IFNγ、N-IL-2的重组质粒。这些重组质粒在真核细胞中得到了表达。用这些真核表达质粒可以免疫小猪,通过检测其对猪体免疫应答的诱导能力和接种后动物对PRRSV的抵抗力来对基因疫苗的效能进行研究,以期筛选到有效的基因疫苗并为今后进一步的研究提供思路和理论基础。     

小反刍兽疫病毒M基因截短表达及鉴定

目的将小反刍兽疫病毒M蛋白基因截短表达后用于特异性单抗制备及临床抗体水平检测。方法:用在线分析软件BepiPred分析小反刍兽疫病毒M蛋白潜在的B细胞线性表位,以本实验室构建的pCR2.1T-PPRV M质粒为模板,扩增三段截短的M基因,纯化后的PCR产物分别与克隆载体pCR2.1T连接,筛选出的阳性重组质粒经双酶切后,分别与表达载体pET-32a(+)及pGEX-6p-1连接,再将鉴定为阳性的重组质粒转化入E.coli BL21(DE3)菌株诱导表达,并用SDS-PAGE及Western blot验证。结果 PCR产物电泳,得到与预期大小相符的特异性片段。对连接克隆载体及表达载体的重组质粒双酶切后,均出现与预期一致的片段,DNA测序表明,插入片段序列与小反刍兽疫Nigeria75/1株M蛋白基因完全一致。重组蛋白经SDS-PAGE及Western blot鉴定,证明所构建的重组蛋白均获得高效表达,并均具有良好的反应原性。结论成功表达了小反刍兽疫截短M基因的蛋白,为制备特异性单抗及小反刍兽疫抗体检测奠定了一定基础。     

乳牛口服过量维生素A对维生素D3及钙磷代谢的影响

从北京郊区某农场选取１０头健康成年黑白花乳牛，随机分为试验（５头，每周每头口服维生素Ａ１００万单位）和对照（５头，不作任何处理）两组。８周以后检测发现，试验组血清钙和无机磷水平呈下降趋势；尾椎骨皮质变薄，密度下降，呈现骨质吸收现象；血清碱性磷酸酶显著下降（Ｐ＜０．０５）；其活性代谢产物２５－羟维生素Ｄ３（２５－ＯＨ－Ｄ３）极显著下降（Ｐ＜０．０１）；１，２５－二羟维生素Ｄ３［１，２５－（ＯＨ）２－     

中草药与寡糖配伍对肉鸡抗氧化功能的影响

试验研究了中草药女贞子、五味子以及女贞子、五味子与葡萄糖甘露寡糖(MOS)配伍对肉鸡抗氧化指标的影响。选择180只1日龄肉鸡公雏随机分为6组,对照组饲喂基础饲粮,抗生素组在基础饲粮中添加5mg/kg黄霉素,试验组分别在基础饲粮中添加1%女贞子、1%五味子、1%女贞子+0.5%MOS、1%五味子+0.5%MOS。试验从1日龄开始,试验期56d。结果显示:女贞子、五味子可提高肉鸡心脏超氧化物歧化酶(SOD)活性(P<0.05),以及血清和组织谷胱甘肽还原酶(GR)活性(P<0.05),降低血清和心脏丙二醛(MDA)含量(P<0.05)。上述试验结果表明中草药添加剂能够提高肉鸡抗氧化功能。     

A novel dendrimeric peptide induces high level neutralizing antibodies against classical swine fever virus in rabbits

The amino acid sequence (TAVSPTTLR, 829-837aa) on the glycoprotein E2 of classical swine fever virus (CSFV) is a conserved and linear neutralizing epitope. In the present study, two peptides were constructed based the core sequence of this neutralizing epitope, the dendrimeric peptide (Th-B(4)) containing four copies of B cell epitope fused to one copy of promiscuous T helper (Th) cell epitope and the peptide Th-B containing a single copy of B cell epitope fused to one copy of Th cell epitope. The dendrimeric peptide Th-B(4) elicited high titers of neutralizing antibodies as detected in an indirect ELISA, blocking ELISA and neutralization test and induced a complete protection against CSFV C strain in rabbits. The Th-B elicited low titers of neutralizing antibodies and did not induce a protection in rabbits. These results suggest that the dendrimeric peptide Th-B(4) may be a promising marker vaccine candidate against CSFV and the multimerization is a requirement for development of a peptide vaccine.     

赴美采购流产大豆价格变数加大

2009年5月27日,商务部组团赴美采购2000万吨大豆的“超级订单”,最终流产。双方没有像往年一样签订协议,一个重要原因是中方的报价没有得到美方的认同。而幕后的原因是,美方忌惮我国购买大豆用于收储,调控市场价格。     

Bactrian camel (Camelus bactrianus) integrins αvβ3 and αvβ6 as FMDV receptors: Molecular cloning, sequence analysis and comparison with other species

Integrins are heterodimeric adhesion receptors that participate in a variety of cell–cell and cell–extracellular matrix protein interactions. Many integrins recognize RGD sequences displayed on extracellular matrix proteins and the exposed loops of viral capsid proteins. Four members of the αv integrin family of cellular receptors, αvβ3, αvβ6, αvβ1 and αvβ8, have been identified as receptors for foot-and-mouth disease virus (FMDV) in vitro, and integrins are believed to be the receptors used to target epithelial cells in the infected animals. To analyse the roles of the αv integrins from a susceptible species as viral receptors, we have cloned Bactrian camel αv, β3 and β6 integrin cDNAs and compared them to those of other species. The coding sequences for Bactrian camel integrin αv, β3 and β6 were found to be 3165, 2289 and 2367 nucleotides in length, encoding 1054, 762 and 788 amino acids, respectively. The Bactrian camel αv, β3 and β6 subunits share many structural features with homologues of other species, including the ligand binding domain and cysteine-rich region. Phylogenetic trees and similarity analyses showed the close relationships of integrin genes from Bactrian camels, pigs and cattle, which are each susceptible to FMDV infection, that were distinct from the orders Rodentia, Primates, Perissodactyla, Carnivora, Galliformes and Xenopus. We postulate that host tropism of FMDV may in part be related to the divergence in integrin subunits among different species.     

日粮营养对动物脂肪组织基因表达的影响

脂肪组织是一个强大的分泌器官,它能分泌许多与能量、脂肪代谢相关的酶、激素等.调控能量代谢和脂肪组织生长.本文综述了日粮对动物脂肪组织中基因表达作用的研究状况.并阐述了营养成分对这些基因上下游表达的作用机制.     

鸡微卫星DNA指纹与屠宰性状关系的研究

利用25个微卫星标记,对以隐性白羽鸡和仙居鸡为亲本建立的资源群体F2500只鸡进行遗传检测,并测定各个体的活重、屠体重、屠体率,采用方差分析法对标记与性状进行连锁分析.研究结果表明各位点平均等位基因数为5,平均杂合度为0.7012,平均多态信息含量为0.6468.方差分析显示MCW0095、ABR0322、ADL289、MCW4、ADL166、MCW104对活重、屠体重有显著影响(P＜0.05或0.01);MCW104对屠体率有极显著影响(P＜0.01);其他标记对性状的影响未达到显著水平.     

Marek's disease resistant/susceptible MHC haplotypes in Xiayan chickens identified on the basis of BLB2 PCR-RFLP and BLB2/BF2 sequence analyses

1. The aim was to analyse the variability of the BLB2/BF2 genes of Xiayan chickens to identify homozygous birds with resistance or susceptibility to Marek's disease (MD).

2. The experiment used two lines: birds from a common line were divided into Group A (unvaccinated) and Group B (vaccinated with herpesvirus of turkeys (HVT)); and birds from an MD-resistant line were divided into Group C (unvaccinated) and Group D (vaccinated with HVT). They were challenged intra-abdominally with Marek's disease virus (MDV) and genomic DNA extracted from peripheral blood lymphocytes so that polymorphism of the BLB2/BF2 genes could be analysed using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) and nucleotide sequence analysis.

3. A 374-bp fragment of the BLB2 gene was amplified from the samples and, after digesting with restriction enzymes Alu I, Cai I, Cfr I, Hin1 I, Hinf I and Rsa I for RFLP analysis, the 6 electrophoretic patterns were analysed. Seven homozygous genotypes were found and used tentatively to identify alleles of the BLB2 gene.

4. A 765-bp fragment of the BF2 gene was amplified from the 7 samples for cloning and sequencing.

5. Six homozygous birds were confirmed from the sequenced BLB2/BF2 gene. Four birds were resistant to MD. Three birds had identical nucleotide sequences and were highly homologous with MHC haplotype B⁶, which is MD resistant. One bird had high homology with the highly MD-resistant B²¹ haplotype, and two birds were susceptible and highly homologous to the B¹⁹ haplotype, which is highly MD susceptible.