马立克氏病病毒超强毒感染鸡羽髓蛋白质组分析

【目的】羽毛是细胞游离马立克氏病病毒(Marek’s disease virus,MDV)释放的部位,为了解感染MDV后鸡羽中宿主基因表达的变化及对病毒感染的应答,进行了MDV感染鸡的羽髓蛋白质组学分析。【方法】1日龄无特定病原体(specific pathogen free,SPF)鸡人工感染MDV超强毒RB1B株(1000PFU),感染后21d采集鸡羽毛,提取羽髓蛋白,以17cm,pH5-8的IPG胶条进行二维电泳,以未感染病毒的SPF鸡羽髓蛋白为对照,使用PDQuest软件对二维电泳图谱进行差异蛋白分析,并选取部分差异斑点进行质谱鉴定。【结果】PDQuest软件分析发现攻毒组和对照组表达差异大于两倍的蛋白点有41个,其中攻毒组表达上调的蛋白点25个,下调的蛋白点7个,新出现的蛋白点有9个。质谱分析共成功鉴定了21个斑点,对应于20个蛋白。如载脂蛋白AI(apolipoprotein AI)、14-3-3 sigma(两个斑点均为该蛋白)、癌蛋白18(stathmin)等。【结论】功能预测表明这些蛋白涉及到宿主的抗病毒应答、物质代谢、细胞骨架成分、细胞增殖相关等方面。     

球囊菌胁迫中华蜜蜂幼虫肠道过程中病原的转录组学研究

【目的】本研究利用RNA-seq技术对球囊菌胁迫的中华蜜蜂(中蜂)幼虫肠道进行深度测序,经趋势分析得到差异表达基因(DEGs)的显著表达模式,进而对胁迫过程中的球囊菌进行转录组学分析。【方法】利用Illumina HiSeq 2500平台对球囊菌胁迫的中蜂幼虫肠道进行深度测序,并利用相关软件进行了深入分析。最后,通过RT-qPCR对RNA-seq数据进行了验证。【结果】本研究共得到球囊菌的41133932条高质量clean reads。22865个DEGs共聚类为8个基因表达模式,其中,16769个DEGs聚类为2个显著上调趋势与2个显著下调趋势。GO富集分析结果显示,显著上调与显著下调趋势中的DEGs分别富集于40与37个GO term,基因富集数最多的为细胞进程(2486 unigenes)。KEGG代谢通路(pathway)富集分析结果显示,显著上调与显著下调趋势中的DEGs分别富集于119和112个pathway,基因富集数最多的分别是氨基酸生物合成(127 unigenes)与核糖体(98 unigenes)。进一步分析表明球囊菌在胁迫中蜂幼虫肠道的过程中通过提高物质合成促进其增殖,而宿主通过抑制球囊菌的蛋白合成抵御病原入侵。富集在MAPK信号通路的11个DEGs的表达水平随着胁迫时间的延长而逐渐下降,推测中蜂幼虫通过抑制该通路而阻遏球囊菌增殖。【结论】本研究不仅为揭示白垩病过程中的球囊菌-中蜂幼虫互作提供了重要信息,也为阐明不同抗性蜂种的球囊菌抗性差异奠定了基础。     

有机配体对稀土元素在小麦体内积累和分异的影响

采用营养液培养和添加外源混合稀土等方法,研究了有机配体柠檬酸、EDTA和DTPA对稀土元素在小麦的根和叶中积累与分异的影响。结果表明,低浓度有机配体对小麦根和叶中的稀土元素,尤其是轻稀土元素的积累有轻微的促进作用,随浓度的升高则表现出显著的降低作用。有机配体对重稀土的作用比轻稀土强,使根和叶中稀土元素的分布曲线向重稀土相对亏缺的方向发展。3种配体对轻、重稀土分异的作用强度为:EDTA>DTPA>柠檬酸。通过VM INTEQ计算表明,在EDTA和DTPA作用下小麦叶中稀土元素的积累与轻、重稀土的分异主要由溶液中呈自由离子态稀土元素的含量和组成控制;柠檬酸作用下小麦叶中稀土元素的变化与自由离子态稀土的含量和组成关系较弱,推测REE-柠檬酸络合物可被小麦直接吸收并运转到小麦的叶中。     

Protein kinase C delta null mice exhibit structural alterations in articular surface,intra-articular and subchondral compartments

IntroductionStructural alterations in intra-articular and subchondral compartments are hallmarks of osteoarthritis, a degenerative disease that causes pain and disability in the aging population. Protein kinase C delta (PKC-δ) plays versatile functions in cell growth and differentiation, but its role in the articular cartilage and subchondral bone is not known.MethodsHistological analysis including alcian blue, safranin O staining and fluorochrome labeling were used to reveal structural alterations at the articular cartilage surface and bone–cartilage interface in PKC-δ knockout (KO) mice. The morphology and organization of chondrocytes were studied using confocal microscopy. Glycosaminoglycan content was studied by micromass culture of chondrocytes of PKC-δ KO mice.ResultsWe uncovered atypical structural demarcation between articular cartilage and subchondral bone of PKC-δ KO mice. Histology analyses revealed a thickening of the articular cartilage and calcified bone–cartilage interface, and decreased safranin O staining accompanied by an increase in the number of hypertrophic chondrocytes in the articular cartilage of PKC-δ KO mice. Interestingly, loss of demarcation between articular cartilage and bone was concomitant with irregular chondrocyte morphology and arrangement. Consistently, in vivo calcein labeling assay showed an increased intensity of calcein labeling in the interface of the growth plate and metaphysis in PKC-δ KO mice. Furthermore, in vitro culture of chondrocyte micromass showed a decreased alcian blue staining of chondrocyte micromass in the PKC-δ KO mice, indicative of a reduced level of glycosaminoglycan production.ConclusionsOur data imply a role for PKC-δ in the osteochondral plasticity of the interface between articular cartilage and the osteochondral junction.

Electronic supplementary material

The online version of this article (doi:10.1186/s13075-015-0720-4) contains supplementary material, which is available to authorized users.     

新城疫病毒非编码序列的遗传演化趋势

【目的】探讨新城疫病毒全基因序列中非编码序列的分子演化规律。【方法】结合本研究室2012年自产蛋下降鸭群中分离测序的一株鸭源新城疫病毒全序列,从GenBank下载35株不同基因型新城疫病毒全长cDNA序列,获取非编码序列,分别绘制引导序列、尾随序列、F-HN及HN-L基因间隔序列(IGS)的遗传进化树,比较编码基因内5’及3’UTR序列核苷酸序列替代特点。【结果】非编码序列的长度及位置高度保守,而其核苷酸基因序列在不断发生变异,且变异趋势与编码基因序列相一致。【结论】新城疫病毒在整个基因组上编码和非编码序列同步发生变异。     

Kosteletzkya virginica, a halophytic species with potential for agroecotechnology in Jiangsu Province, China

Kosteletzkya virginica (L.) Presl. is a perennial dicot halophytic species, that grows in brackish portions of coastal tidal marshes of the mid-Atlantic and southeastern United States. New saline mudflats have been increasing every year in Northern Jiangsu, China. In 1993, we introduced K. virginica (L.) into China from the Halophyte Biotechnology Center (University of Delaware, USA) as a potential species to improve the soil and develop ecologically sound saline agriculture. Our nearly 10 years of experimental research, both in a test garden and in the field, indicated that K. virginica adapts excellently to the heavy saline soils of Jiangsu Province, China. The average seed yield of K. virginica in 2001 at Yancheng was 638 kg/ha from 1-year old seedlings. There were significant variations among individuals from the unselected population of K. virginica on growth, quality, and seed yield traits. There is great potential to increase the seed yield if superior clones are selected. The seed yield of 35 selected individuals was six times greater than that of the average. Four growth traits of K. virginica were found to have a significant correlation with seed yield. However, there was no strong positive or negative correlation among seed quality traits. The mean seed weight and the germination ratio of the 35 selected individuals of K. virginica were 16.36±0.32 g and 76.11±1.82%, respectively. The percentage content of oil and crude protein in the seeds was 11.28±0.67 and 8.17±0.19%, respectively. The percentage of seed vigor of the selected individuals was 99%, as determined by soft X-ray radiography.     

利用土壤宏基因组技术定向筛选抗生素产生菌的方法学建立

为了快速从土壤中定向筛选抗生素产生菌, 本研究利用宏基因组技术进行了土壤中抗生素产生菌快速筛选方法的探索。对6 种不同土质的土壤利用液氮冷冻法提取土壤基因组DNA 并用PVPP柱层析法进行纯化, 每克湿土可得到32.25?61.25 μg DNA, 片段大小为23 kb左右, 说明宏基因组DNA样品完整, 16S rDNA的 PCR结果证明了该基因组DNA纯度可用于后期的分子操作。利用Herbimycin产生菌掺入法进行了抗生素产生菌筛选方法学的验证。结果证明, 每克湿土掺入103个Herbimycin产生菌时, 利用Herbimycin合成基因簇的特定引物即可从所提取的土壤基因组DNA中扩增出目标条带, 并且可以利用传统菌种分离方法从土壤中重新分离出来被掺入到土壤的Herbimycin产生菌。这些结果证明本研究建立的定向抗生素产生菌筛选方法, 具有快捷、灵敏的特点, 大大减少传统筛选方法的工作强度和时间, 为微生物新药的研发建立一种全新的研究方法。     

芒颖大麦草对菱镁矿粉尘污染的生态适应性

采用盆栽试验,向土壤表面投加不同量(0、1％、5％、10％、15％、20％、30％)的菱镁矿粉尘,研究土壤理化性质和芒颖大麦草对镁的耐性.结果表明:菱镁矿粉尘施加到土壤表面时,可快速形成结皮.当菱镁粉尘比例超过20％时,土壤理化性质发生明显变化,表现为pH显著升高,电导率逐渐降低,有机质含量先降低后升高;芒颖大麦草的叶绿素含量随着镁粉尘含量的增加呈现先增加后降低的趋势,而可溶性蛋白含量随着镁粉尘含量的增加呈现先增加后降低再升高的趋势;芒颖大麦草具有较强的耐镁特性,土壤中镁浓度达到4.61 g·kg-1时,芒颖大麦草仍生长良好.芒颖大麦草适宜作为菱镁矿区撂荒地生态恢复的植物种.     

Surface Display of Human Serum Albumin on Bacillus subtilis Spores for Oral Administration

Human serum albumin (HSA) is the major protein component of human plasma. To date, HSA for clinical uses is mostly produced by fractionation of human whole blood, which is accompanied by a lot of limitations. To obtain long-term bioactive albumin, we used hsa as a foreign gene and constructed a recombinant plasmid pJS700-HSA which carries a recombinant gene cotC-hsa under the control of cotC promoter. Plasmid pJS700-HSA was transformed into Bacillus subtilis by double cross-over and an amylase inactivated mutant was produced. After induction of spore formation, western blot and fluorescence immunoassay were used to monitor HSA surface expression on spores. We estimated that HSA displayed on the spore accounted for 0.135 % of the total spore proteins and about 0.023 fg HSA were exposed on the surface of each spore. Oral administration to mice with spores displaying HSA implied that the recombinant spores may have potential ability to increase the serum albumin level in vivo due to the resistant characters of spores.