HIV-1 Env三聚体抗原改造研究进展

HIV-1疫苗研发是当前艾滋病研究的一大热点,其病毒表面包膜糖蛋白Env三聚体介导病毒与细胞融合,是HIV-1疫苗研究的重要靶点。因此,设计并在体外表达类天然Env三聚体对HIV-1疫苗的研发具有重要的意义。近年来,Env三聚体研究取得了显著的进展。SOSIP、NFL2P、UFO等抗原改造方法实现了类天然Env三聚体的体外表达,逐步解决了改造抗原产量低、结构不稳定等问题,且表达的Env三聚体抗原能在动物免疫中诱导机体产生较高的中和抗体水平。Env三聚体改造方法促进了HIV-1疫苗的研究。文中综述了SOSIP、NFL2P、UFO三种HIV-1 Env三聚体抗原改造方法,对比各个改造方法优缺点,并结合自身工作提出相关建议,为后续HIV-1抗原的相关设计提供指导。     

A 3.1-kb genomic fragment of Bacillus subtilis encodes the protein inhibiting growth of Xanthomonas oryzae pv. oryzae

AIMS: To clone genes of Bacillus subtilis encoding peptides that inhibit the growth of Xanthomonas orzae pv. oryzae (Xoo). METHODS AND RESULTS: A 3.1-kb DNA fragment from B. subtilis SO113 encoding peptides that inhibit the growth of Xoo (anti-Xoo, showing an inhibition zone) was isolated from a plasmid library of B. subtilis 6 GM15. Sequence analysis revealed that it contained three complete open reading frames (ORFs): ybcO, ybcS and a novel ORF designated ybcPQ. Deleting the last 96 bp of ybcS from the plasmid eliminated the anti-Xoo activity, suggesting that ybcS is required for producing the anti-Xoo activity. However, no anti-Xoo activity could be detected for the plasmid with ybcS alone. Further analysis showed that ybcO, at least, was also required to obtain the anti-Xoo activity. CONCLUSIONS: A fragment of B. subtilis has been cloned that expresses an anti-Xoo activity that requires ybcS and ybcO. SIGNIFICANCE AND IMPACT OF THE STUDY: These genes could be useful for the genetic engineering of resistance to rice bacterial diseases and for the design of new anti-Xoo biocontrol agents.     

半夏属 (PINELLIA)的花粉粒和醇溶蛋白比较

继细胞学 〔1〕、形态学 〔2〕和比较解剖学 〔3〕的研究之后 ,我们又对半夏属的花粉粒和醇溶蛋白作了比较分析 ,旨在探讨这些性状的变异与分类学价值。1 　材料和方法1 .1　实验材料　实验材料采自华东 3省的不同地区。在南京栽植至少 1年后 ,在盛花季节 ( 5月 )选有花植株的花粉和地下块茎作实验分析。各材料的种名、群体编号和产地见表 1。表 1 　供实验用的半夏属 5种植物群体材料的来源Table1　 Sources of material for experiment of the populationsbelonging to5species of Pinellia种名Species群体编号No.of populations产地Lo…     

Projecting future grassland productivity to assess the sustainability of potential biofuel feedstock areas in the Greater Platte River Basin

This study projects future (e.g., 2050 and 2099) grassland productivities in the Greater Platte River Basin (GPRB) using ecosystem performance (EP, a surrogate for measuring ecosystem productivity) models and future climate projections. The EP models developed from a previous study were based on the satellite vegetation index, site geophysical and biophysical features, and weather and climate drivers. The future climate data used in this study were derived from the National Center for Atmospheric Research Community Climate System Model 3.0 ‘SRES A1B’ (a ‘middle’ emissions path). The main objective of this study is to assess the future sustainability of the potential biofuel feedstock areas identified in a previous study. Results show that the potential biofuel feedstock areas (the more mesic eastern part of the GPRB) will remain productive (i.e., aboveground grassland biomass productivity >2750 kg ha^?1 year^?1) with a slight increasing trend in the future. The spatially averaged EPs for these areas are 3519, 3432, 3557, 3605, 3752, and 3583 kg ha^?1 year^?1 for current site potential (2000–2008 average), 2020, 2030, 2040, 2050, and 2099, respectively. Therefore, the identified potential biofuel feedstock areas will likely continue to be sustainable for future biofuel development. On the other hand, grasslands identified as having no biofuel potential in the drier western part of the GPRB would be expected to stay unproductive in the future (spatially averaged EPs are 1822, 1691, 1896, 2306, 1994, and 2169 kg ha^?1 year^?1 for site potential, 2020, 2030, 2040, 2050, and 2099). These areas should continue to be unsuitable for biofuel feedstock development in the future. These future grassland productivity estimation maps can help land managers to understand and adapt to the expected changes in future EP in the GPRB and to assess the future sustainability and feasibility of potential biofuel feedstock areas.     

戊型肝炎病毒荧光定量RT-PCR快速检测技术的建立和初步应用

利用DNAMAN软件对GenBank登录的戊型肝炎病毒四个主要基因型代表株的序列进行分析, 选择其高度保守的ORF2区域设计合成引物和探针, 并用包含有扩增区域的核苷酸片段进行体外转录制备标准品cRNA。在对荧光定量RT-PCR的反应条件优化的基础上, 建立了适用于戊型肝炎病毒主要基因型检测的荧光定量RT-PCR检测技术。该检测技术可以有效检测I型和IV型戊型肝炎阳性病料, 而对猪的其它几种疫病阳性病料则为阴性结果, 证实本技术的特异性强、可靠性好。对阳性标准品的检测结果表明, 所建立的TaqMan荧光定量RT-PCR灵敏度可达2.0×101拷贝/反应, 相比于巢式RT-PCR方法, 其灵敏度高10~100倍以上。在对54份临床样品的检测中, 进一步证实了该方法快速、灵敏且重复性好, 可满足戊型肝炎病毒早期快速诊断的需要。     

海枣曲霉β—D—岩藻糖苷酶的研究

Although beta-D-fucosidase (beta-D-fucoside fucohydrolase, EC 3.2.1.38) has been isolated from various sources, the identity of this enzyme is still not settled. We have purified a specific beta-D-fucosidase in electrophoretically homogeneous form crude extracts of Aspergillus phoenicis by polyethyleneglycol 6000-phosphate buffer aqueous two-phase separation, and successive chromatography on DEAE-Sephadex A-50, hydroxyapatite and Sephadex G-100 columns. The molecular weight of the enzyme was estimated to be 57000 by SDS-polyacrylamide gel electrophoresis and 50000 to 60000 by gel filtration on Sephadex G-100. The enzyme showed optimum coside were 2.4mmol/L, and 1.28 mumol min-1 the pH range 5.5-6.5 and below 35 degrees C. The Km and the Vmax values for pNP-beta-D-fucoside were 2.4mmol/L, and 1.28 mumol.min-1.mg-1 respectively. The enzyme was strongly inhibited by sulfhydryl group reagents, PCMB-NEM and iodoacetate. It was also inhibited by EDC, DEP and NBS. Thus, -SH, -COOH groups, histidyl and tryptophyl residues were essential for enzyme activity. The purified beta-D-fucosidase showed high specificity toward p-nitrophenyl beta-D-fucoside. The enzyme was inhibited by D-fucose and D-fucono-gamma-lactone, but not by D-galactose, D-galactono-gamma-lactone, D-glucose or D-glucono-gamma-lactone; the latter compounds are specific inhibitors of beta-D-galactosidase and beta-D-glucosidase respectively. Thus, this enzyme is the most strictly specific beta-D-fucosidase when compared with those previously reported.     

Analysis of synonymous codon usage in H5N1 virus and other influenza A viruses

In this study, we calculated the codon usage bias in H5N1 virus and performed a comparative analysis of synonymous codon usage patterns in H5N1 virus, five other evolutionary related influenza A viruses and a influenza B virus. Codon usage bias in H5N1 genome is a little slight, which is mainly determined by the base compositions on the third codon position. By comparing synonymous codon usage patterns in different viruses, we observed that the codon usage pattern of H5N1 virus is similar with other influenza A viruses, but not influenza B virus, and the synonymous codon usage in influenza A virus genes is phylogenetically conservative, but not strain-specific. Synonymous codon usage in genes encoded by different influenza A viruses is genus conservative. Compositional constraints could explain most of the variation of synonymous codon usage among these virus genes, while gene function is also correlated to synonymous codon usages to a certain extent. However, translational selection and gene length have no effect on the variations of synonymous codon usage in these virus genes.     

Cellular Source and Mechanisms of High Transcriptome Complexity in the Mammalian Testis

1. Download : Download full-size image

     

Development of polymorphic microsatellite markers in Camellia chekiangoleosa (Theaceae) using 454-ESTs

? Premise of the study: A set of microsatellite markers for Camellia chekiangoleosa was developed and characterized using 454 sequencing technology to study the population genetic structure and the diversity of germplasm collections. ? Methods and Results: Eighteen polymorphic microsatellite markers were identified and tested in 150 individuals from three natural populations of C. chekiangoleosa. Alleles numbered from two to seven, and the observed and expected heterozygosities ranged from 0.100 to 0.760 and 0.133 to 0.809, respectively. ? Conclusions: These markers will potentially be conducive to further genetic studies on C. chekiangoleosa.     

Comparison of sex ratio and cell number of IVM-IVF bovine blastocysts co-cultured with bovine oviduct epithelial cells or with Vero cells

The influence of 2 co-culture systems (BOEC and Vero cells) on the development rates, quality grades and sex ratios of IVM-IVF bovine embryos were studied. Zygotes obtained after IVF were co-cultured in each co-culture system for 7 and 8 d (Day 0 = day of insemination) in B2 medium. No effect of the co-culture system was observed on development rates measured on Days 7 and 8. However, Vero cell co-culture had a positive influence on embryo quality. Irrespective of their sex, embryos produced on Vero cells showed higher cells number than those co-cultured on BOEC (103.4 +/- 3.8 and 97 +/- 8.12 for BOEC vs 113.7 +/- 3.5 and 114 +/- 5.9 for Vero cells at Days 7 and 8, respectively; P < 0.05). The percentage of male embryos was increased in the two co-culture systems (60.7% males for BOEC; P < 0.05 vs 63% males for Vero cells; P < 0.01) on Day 7. In both co-culture systems the increase in the percentage of males was more obvious for embryos reaching the most advanced stage (expanded blastocysts). The results show that Vero cells improved the quality grade of bovine embryos produced in vitro, and thus are recommended for use as a safe co-culture system that does not contain pathogens.