A dominant antigenic epitope on SARS-CoV spike protein identified by an avian single-chain variable fragment (scFv)-expressing phage

Severe acute respiratory syndrome (SARS) is a newly emergent human disease, which requires rapid diagnosis and effective therapy. Among antibody sources, immunoglobulin Y (IgY) is the major antibody found in chicken eggs and can be used as an alternative to mammalian antibodies normally used in research and immunotherapy. In this study, phage-expressing chicken monoclonal scFv antibody was chosen and characterized with phage display antibody technology. Truncated fragments of SARS-CoV spike protein were cloned in pET-21 vector and expressed in BL-21 Escherichia coli (E. coli) cells. After purification, the purity of these recombinant spike proteins was examined on SDS-PAGE and their identity verified with Western blot analysis using anti-his antibodies and sera from convalescent stage SARS-CoV-infected patients. Using these bacteria-derived proteins to immunize chickens, it was found that polyclonal IgY antibodies in the egg yolk and sera were highly reactive to the immunogens, as shown by Western blot and immunocytochemical staining analysis. A phage displaying scFv library was also established from spleen B cells of immunized chicken with 5 x 10(7) clones. After four panning cycles, the eluted phage titer showed a 10-fold increase. In sequence analysis with chicken germline gene, five phage clones reacted, with large dissimilarities of between 31 and 62%, in the complementarity-determining regions, one dominant phage 4S1 had strong binding to fragment Se-e, located between amino acid residues 456-650 of the spike protein and this particular phage had significantly strong binding to SARS-CoV-infected Vero E6 cells. Based on the results, we conclude that generating specific scFv-expressing phage binders with the phage display system can be successfully achieved and that this knowledge can be applied in clinical or academic research.     

不同的干燥工艺对荷花蜂花粉功能成分的影响

探讨了5种干燥工艺对荷花蜂花粉功能成分的影响。微波干燥的干燥效率最高,60%火力处理540s可使水分从31.41%降到6.42%;干燥方式对荷花蜂花粉蛋白质影响较小;微波干燥后总黄酮提取率最高,为13.20mg/g;真空冷冻干燥后维生素C和过氧化氢酶含量最高,分别为48.89mg/100g和29.86mg/g·min。总的来说,真空冷冻干燥对荷花蜂花粉的功能成分影响最小;常压恒温干燥对功能成分影响较小、工艺简单、对设备要求低,宜于大规模应用于实际生产中。     

重要营养素--血红素铁

血红素铁是卟啉类化合物,是哺乳动物血液中的天然色素和生物铁,广泛应用于医药、食品和化工业。文章就血红素铁的理化特性、生理功能及应用进行了简述,并对其常用提取方法进行了比较概述。     

草地畜牧业是推动黔西南州农业结构战略牲调整的突破口

改革开放20多年来。黔西南州农业获得了长足发展，在产品供求关系、生产目标、增长方式、经营形式、产业关联等方面都发生了显著变化，进入了农产品供求基本平衡，以提高品质、优化结构和增加农民收入为主，注重传统投入与资本集约和技术集约相结合的优化发展阶段。加入WTO，对于我州正处在发展新阶段的农业和农村经济来说。是一次重要的机遇和挑战。面对激烈的国际、国内市场竞争。在西部大开发中，为了改善生态环境，调整农业结构，增加农民收入，州委、州政府决定把草地畜牧业作为我州农业和农村产业结构调整的重点。制定了百万亩草地畜牧业发展规划。出台了加快草地畜牧业发展的实施意见。     

猪毛滴虫病病原及诊治的研究进展

猪毛滴虫病的历史较短,关于该病的研究和病例报道也较少。近年来,随着该病导致保育猪腹泻病例日益增多,猪毛滴虫病对养猪业的危害日益显现。本文就该病的病原、危害性、致病机理、流行病学、临床症状、病理变化、诊断以及防治作一综述,旨在为猪毛滴虫病的研究和防控提供参考。     

益生菌发酵饲料对猪的生长和肉质的影响

应用益生菌发酵饲料喂猪,能提高饲料中粗蛋白含量,明显降低猪粪臭味,促进生长、降低料肉比和提高猪肉品质。     

Gene cloning of porcine adiponectin gene from adipose tissue and construction of its eukaryotic expression vector

To clone adiponectin (ADPN) gene from Shaziling porcine adipocyte and construct its eukaryotic expression vector, total RNA was extracted from subcutaneous fatty tissue. One pair of specific primers was designed by Primer 5.0 software according to the sequence of ADPN gene of porcine available in GenBank. The ADPN gene was amplified by PCR from cDNA and cloned into pMD18‐T vector to construct recombinant clonal vector pMD‐ADPN, sequenced and analysed. A recombinant expression plasmid pPICZaA‐ADPN was constructed by subcloning the cloned ADPN gene into the linearized pPICZaA vector. Then, the plasmid pPICZaA‐ADPN was expressed in Pichia pastoris (GS115) by electrotransformation. Western blot and Bradford analysis were used to determine the target protein induced by methanol. Results showed that the genome size of ADPN was 732 bp and encoded 244 amino acid, the nucleotide sequence of ADPN shared 100% identity with that of porcine available in GenBank. Western blot and Bradford analysis showed that the recombinant ADPN was expressed in GS115 correctly and has certain immune activity. The expression level of ADPN was 28.5 μg/ml. In conclusion, the recombinant ADPN could express in eukaryotic expression vector pPICZaA‐ADPN constructed in this study effectively.     

如何科学寄养初生仔犬

一、采取寄养措施的前提1.母犬产后死亡。2.母犬产后乳汁分泌过少,哺育较多的仔犬有困难。3.母犬产仔过多,超过其有效哺乳头数。4.不同品种仔犬的经济价值不同,利用奶水较好的母犬来哺育经济价值高的仔犬。二、寄养方法主要有直接寄养和交叉寄养两种方法。1.直接寄养是指直接把     

法国犬协印象

2006年8月23日,有幸随公安部警犬技术培训团赴欧洲考察培训。此次培训范围广、安排紧、内容多。根据日程安排,8月28日我们考察了有100多年历史的法国养犬协会(SociétéCentraleCanine,SCC)。法国养犬协会(SCC)总部位于法国首都巴黎,成立于1882年,其前身是法国的一个猎犬俱乐部     

Interpopulation and intrapopulation maternal lineage genetics of the Lanyu pig (Sus scrofa) by analysis of mitochondrial cytochrome b and control region sequences

The Lanyu pig is an indigenous breed from the Lanyu Islet, which is southeast of Taiwan. Two herds of Lanyu pigs were introduced from the Lanyu Islet into Taiwan in 1975 and 1980. The current population of conserved Lanyu pigs consists of only 44 animals with unknown genetic lineage. The Lanyu pig possesses a distinct maternal genetic lineage remote from Asian and European pigs. The present study aimed to understand the phylogenetic relationship among conserved Lanyu, Asian, and European type pigs based on the cytochrome b coding gene, to ascertain the maternal lineage and genetic diversity within the conserved Lanyu pigs, and to address whether genetic introgression from exotic or Formosan wild pigs had occurred in the conserved Lanyu pigs. Entire mitochondrial genomes of both types of Lanyu pig comprised 2 ribosomal RNA, 22 transfer RNA, and 13 protein-coding genes. Only 2 haplotypes of the mitochondrial DNA (mtDNA) control region and cytochrome b were identified in the conserved Lanyu pig herds. When maximum likelihood trees were constructed, the Type I Lanyu mitochondrial genes formed a unique clade with a large pairwise distance of both cytochrome b and the control region from Asian and European type breeds, Formosan wild pigs, and exotic breeds. Significant loss of genetic diversity of mtDNA within the conserved Lanyu pigs was demonstrated by low haplotype and nucleotide diversities, supported by Fu and Li's D* neutrality test (1.44055; P < 0.05). The mtDNA control region sequences of extant pigs in the Lanyu Islet, however, showed high haplotype and nucleotide diversity, and clustered with exotic pigs. These results indicate no maternal lineage mtD-NA gene introgression from Formosan wild pigs and introduced exotic pigs to conserved Type I Lanyu pigs, and a severe loss of heterozygosity of mtDNA in conserved Lanyu pigs. The remaining extant pigs on the Lanyu Islet have been introgressed with exotic breeds. Strategies for future conservation of native Lanyu pigs are now even more urgent and important.