Comparative Analysis of Eubacterial DNA Polymerase III Alpha Subunits

DNA polymerase III is one of the five eubacterial DNA polymerases that is re-sponsible for the replication of DNA duplex. Among the ten subunits of the DNApolymerase III core enzyme, the alpha subunit catalyzes the reaction for polymer-izing both DNA strands. In this study, we extracted genomic sequences of thealpha subunit from 159 sequenced eubacterial genomes, and carried out sequence-based phylogenetic and structural analyses. We found that all eubacterial genomeshave one or more alpha subunits, which form either homodimers or heterodimers.Phylogenetic and domain structural analyses as well as copy number variations ofthe alpha subunit in each bacterium indicate the classification of alpha subunit intofour basic groups: polC, dnaE1, dnaE2, and dnaE3. This classification is of essencein genome composition analysis. We also consolidated the naming convention toavoid further confusion in gene annotations.     

湖北海棠DNA的CTAB改良法提取及其SSR反应体系优化

为了利用分子标记技术研究湖北海棠的起源、系统发育和分类问题,本研究对湖北海棠及其近缘种叶DNA的CTAB提取法进行改良,并对SSR体系优化.结果表明,将EDTA用量从20 mmol/L提高到80 mmol/L,加大PVP和RNA酶的用量,有助于提高从湖北海棠叶中提取的DNA质量;利用反复实验优化的SSR体系,可从40对引物中筛选出18对多态性好、条带清晰的湖北海棠SSR引物.本研究筛选出的多态引物,将为探讨湖北海棠的起源、进化和遗传变异等提供基础资料.     

谢瓦氏曲霉间型变种YchF基因cDNA全长克隆及序列分析

从谢瓦氏曲霉间型变种抑制性差减杂交(SSH)文库中,筛选到一个子囊孢子时期差异表达的序列标签A0128.860,以其序列为模板设计特异性引物,利用RT-PCR及RACE技术克隆获得了该基因的全长cD-NA,命名为eYchF.生物信息学分析显示,该基因包含一个1182 bp的完整开放阅读框(ORF),编码393个氨基酸,相对分子质量为43.4669 kD,预测的理论等电点为6.99,包括一个保守的G domain (guanine nu-cleotide-binding domain)和一个TGS (ThrRS, GTPase, SpoT) domain,为疏水蛋白.序列同源性分析表明:该基因与棒曲霉(Aspergillus clavatus NRRL 1)的GTP结合蛋白YchF相似度为84%,谢瓦氏曲霉间型变种YchF基因的克隆及生物信息学分析为YchF的进一步研究奠定了基础.     

热带假丝酵母产生子囊孢子的条件及单倍体分离的研究

本文研究了诱导热带假丝酵母子囊孢子产生和子囊破壁的条件,以及单倍体分离和鉴别方法.结果表明:NaAc、KCl这两种成分即可满足热带假丝酵母的产孢营养要求,在含有NaAc 8.2 g/L、KCl 1.8 g/L的琼脂培养基中,28℃培养7 d,热带假丝酵母细胞的产孢率可达到47.5%;单纯使用蜗牛酶对破除热带假丝酵母子囊壁的效率甚低,酶解液加入适量的石英砂同时振荡处理4 h左右,可以显著提高对热带假丝酵母子囊破壁速率.用这种方法处理,绝大多数子囊壁均可破除.子囊破壁处理后再以52℃处理10 min杀死残余的营养体细胞,分离物的单倍体孢子比例可由灭活处理前的48%,提高到76%以上.     

猪瘟病毒NS2-3抗原集中区基因的原核表达及鉴定

为获得猪瘟病毒(classical swine fever virus, CSFV) NS2-3抗原集中区蛋白,并建立CSFV抗体快速检测方法.本研究以CSFV全长基因组质粒为模板,PCR扩增NS2-3抗原表位集中区,利用扩增片段和克隆载体,构建重组表达质粒,命名为pET32a-NS2-3-1.重组表达质粒转化Rosetta (DE3)细胞,利用IPTG诱导表达, SDS-PAGE电泳和Western-blot鉴定重组表达产物.结果表明,重组质粒pET32a-NS2-3-1在28℃诱导5 h得到高效表达,重组蛋白能够与兔抗CSFV阳性血清发生反应.获得CSFV NS2-3抗原集中区蛋白,并且获得的重组蛋白具有抗原性,能够作为CSFV抗体检测的抗原.     

Myogenin蛋白在五指山猪和长白猪骨骼肌中的表达

本研究采用免疫荧光及免疫组化DAB显色技术来检测Myogenin蛋白在五指山猪和长白猪胎儿和成年猪的骨骼肌中的定位和表达情况,并通过平均光密度统计分析Myogenin蛋白在五指山猪和长白猪胎儿和成年猪骨骼肌中表达水平.结果表明:Myogenin蛋白表达定位于骨骼肌细胞核表达;在五指山猪和长白猪胎儿和成年猪的腿部骨骼肌和背部骨骼肌中均检测到阳性产物,在猪胎儿期间骨骼肌中Myogenin蛋白的表达水平高于其在成熟猪骨骼肌中的表达;且在同种猪腿部骨骼肌中的表达水平显著高于其在背部骨骼肌中的表达;此外还检测到Myogenin蛋白在五指山猪骨骼肌中表达均显著高于其在长白猪骨骼肌中的表达(p<0.05).本研究得到的Myogenin蛋白在五指山猪和长白猪骨骼肌中存在显著的表达差异结果,为今后研究不同猪种间骨骼肌发育差异及探讨五指山猪矮小机制和发育机理提供了理论基础.     

金沙江上段浮游藻类和着生藻类群落格局及其与环境因子的关系比较研究

比较河流浮游藻类和着生藻类群落的时空格局及其与环境因子关系的差异,有助于了解两类藻的区别与联系。然而,目前这方面的研究还不多。基于2019年秋季和2020年夏季金沙江上段干流17个样点藻类及水体理化指标的调查数据,分析了不同季节浮游藻类和着生藻类群落结构及其主要环境驱动因子,比较了两类藻的多样性格局及其与环境关系的异同。结果发现,调查河段的浮游藻类和着生藻类均以硅藻为主,其中浮游藻类以极小曲壳藻(Achnanthes minutissima)、钝脆杆藻(Fragilaria capucina)、适中舟形藻(Navicula accomoda)为主要优势种,着生藻类以极小曲壳藻(Achnanthes minutissima)、扁圆卵形藻(Cocconeis placentula)、橄榄绿色异极藻(Gomphonema olivaceum)为主要优势种。浮游藻类和着生藻类秋季平均密度分别为:2.41×10~5个/L、9.43×10~3个/cm~2,均明显高于夏季的平均密度(4.84×10~4个/L、4.84×10~3个/cm~2)。两类藻的群落格局表现出明显的季节变化,但只有着生藻分类单元...     

独龙江河流生态系统健康评价

于2018年10—11月,对独龙江水系干流及支流的河流物理生境、水体理化性质和大型底栖动物进行调查,并根据独龙江自身特点,运用改进的河流健康综合评价指数对该地区生态系统健康状况进行了评价。结果表明:该地区调查样点中85.2%的样点处于健康状况,仅人口密集的乡镇附近健康状况处于亚健康水平,其中旅游业发达的独龙江乡健康状况最差。河流是水资源的重要载体,随着当地旅游业的不断发展,科学评价当地河流的健康状况可以为独龙江河流生态系统修复、水资源的合理开发利用及可持续发展提供重要的科学依据。     

Screening and Assessing 11 Mycobacterium tuberculosis Proteins as Potential Serodiagnostical Markers for Discriminating TB Patients from BCG Vaccinees

Purified protein derivative (PPD) skin tests often yield poor specificity, so that to develop new serological antigens for distinguishing between Mycobacterium tuberculosis infection and Bacille Calmette-Guerin (BCG) vaccination is a priority, especially for developing countries like China. We predicted the antigenicity for selected open reading frames (ORFs) based on the genome sequences of M. tuberculosis H37Rv and M. bovis BCG, as well as their functions and differences of expression under different stim...     

Association of polymorphisms of Nramp1 gene with immune function and production performance of large white pig

The present research was designed to study the association of polymorphism of natural resistance-associated macrophage protein1 (Nramp1) with some immune function and the production performance in Large White pig. The PCR-RFLP technique was applied to analyze the correlation between the polymorphisms of Nramp1 gene and immune function [value of Polymorphonuclear Leukocytes (PMN) obtained by Nitroblue Tetrazolium (NBT) Reduction and effect of Cytotoxin in Monocyte] and production performance in 165 Large White pigs. The results showed that there was one Nde I restriction locus in Large White pig, and both values of PMN by NBT Reduction and effect of Cytotoxin in Monocyte in genotype BB were higher than those in genotype AB (P<0.05). Simultaneously, the weight of 180-day-old pigs with genotype BB was higher than that with genotype AB (P<0.05). The results indicated that there was a significant correlation between different genotypes of Nramp1 gene and Immune function and production performance, and it can be regarded as a candidate gene of disease resistance. All these results provide valuable reference to further studies of pig disease resistance.