从蚕的生命到艺术：中国古代生命观的哲学表达

中国考古出土的蚕业实物及蚕的艺术形象比较丰富,蚕的艺术形象如蚕纹、陶蚕蛹、牙雕蚕、玉石蚕、铜蚕、金蚕等,可统称为"蚕的模拟形态"。对蚕的模拟形态的功用,已有的诸多解释都有待完善。研究表明,蚕的模拟形态或艺术形象表达的功用或为饰品,或为装饰图案,或有待进一步考究。但无论哪种功用,用"蚕"这一形象都蕴含了特有的用意。通过对中国古代生命观的考察,文章认为蚕的艺术形象折射出相应的中国古代哲学生命观,即中国古人追求的死而复生、生生不息、羽化成仙、长乐无极等观念。     

Differential gene expression of the healthy conjunctiva during the day

PurposeTo determine if there is diurnal variation in gene expression in normal healthy conjunctival cells.MethodsBulbar conjunctival swab samples were collected from four healthy subjects in the morning and evening of the same day. The two swab samples were taken from one eye of each participant, with a minimum of five hours gap between the two samples. RNA was extracted and analysed using RNA sequencing (RNA-Seq).ResultsA total of 121 genes were differentially expressed between the morning and the evening conjunctival samples, of which 94 genes were upregulated in the morning, and 27 genes were upregulated in the evening. Many of the genes that were upregulated in the morning were involved in defence, cell turnover and regulation of gene expression, while the genes upregulated in the evening were involved in signalling and mucin production.ConclusionsThis study has identified several genes whose expression changes over the course of the day. Knowledge of diurnal variations of conjunctival gene expression provides an insight into the regulatory status of the healthy eye and provides a baseline for examining changes during ocular surface disease.     

In Vivo Simultaneous Analysis of Gene Expression by Dual-Color Luciferases in Caenorhabditis elegans

Both fluorescent and luminescent observation are widely used to examine real-time gene expression patterns in living organisms. Several fluuorescent and luminescent proteins with specific optical properties have been developed and applied for simultaneous, multi-color observation of more than two gene expression profiles. Compared to fluorescent proteins, however, the application of multi-color luminescent imaging in living organisms is still limited. In this study, we introduced two-color luciferases into the soil nematode C. elegans and performed simultaneous analysis of two gene expression profiles. Using a green-emitting luciferase Eluc (emerald luciferase) and red-emitting luciferase SLR (stable luciferase red), the expression patterns of two genes were simultaneously observed in single animals from embryonic to adult stages over its whole life span. In addition, dual gene activities were observed at the single embryo level, with the simultaneous observation of morphological changes. These are the first application of a two-color luciferase system into a whole animal and suggest that precise relationship of expression patterns of multiple genes of interest can be analyzed over the whole life of the animal, dependent on the changes in genetic and/or environmental conditions.     

Genome-Wide Analyses of the Temperature-Responsive Genetic Loci of the Pectinolytic Plant Pathogenic Pectobacterium atrosepticum

Temperature is one of the critical factors affecting gene expression in bacteria. Despite the general interest in the link between bacterial phenotypes and environmental temperature, little is known about temperature-dependent gene expression in plant pathogenic Pectobacterium atrosepticum, a causative agent of potato blackleg and tuber soft rot worldwide. In this study, twenty-nine P. atrosepticum SCRI1043 thermoregulated genes were identified using Tn5-based transposon mutagenesis coupled with an inducible promotorless gusA gene as a reporter. From the pool of 29 genes, 14 were up-regulated at 18 °C, whereas 15 other genes were up-regulated at 28 °C. Among the thermoregulated loci, genes involved in primary bacterial metabolism, membrane-related proteins, fitness-corresponding factors, and several hypothetical proteins were found. The Tn5 mutants were tested for their pathogenicity in planta and for features that are likely to remain important for the pathogen to succeed in the (plant) environment. Five Tn5 mutants expressed visible phenotypes differentiating these mutants from the phenotype of the SCRI1043 wild-type strain. The gene disruptions in the Tn5 transposon mutants caused alterations in bacterial generation time, ability to form a biofilm, production of lipopolysaccharides, and virulence on potato tuber slices. The consequences of environmental temperature on the ability of P. atrosepticum to cause disease symptoms in potato are discussed.     

计及RSC控制的DFIG短路电流直流分量解析表达

现有文献针对计及转子变流器(RSC)控制的双馈感应风电机组(DFIG)定子短路电流解析表达,将定子磁链当作一阶直流衰减分量或忽略功率外环控制。基于DFIG电压、磁链和RSC控制方程,得到定子电流关于定子电压和定子功率的传递函数,提出定子电流的精确解析表达式。基于RSC内、外环PI参数关系,推导直流分量衰减时间常数和角频率关于PI参数的表达式。分析了RSC内外环PI参数对定子电流直流衰减分量的影响。仿真结果验证了解析表达式的准确性,为PI参数选取和保护装置测量、整定提供依据。     

C4ST-1在大肠杆菌中的可溶性表达

软骨素-4-O-硫酸转移酶-1 (Chondroitin-4-O-sulfotransferase-1,C4ST-1,EC 2.8.2.5)催化软骨素N-乙酰半乳糖胺(N-Acetylgalactosamine,GalNAc)4号位羟基硫酸化生成硫酸软骨素A(chondroitin sulfate A,CSA)。C4ST-1含3对二硫键,在Escherichia coli细胞质内二硫键难以正确形成,故在E. coli中表达时主要以包涵体形式存在。为提高胞内可溶性蛋白质的表达水平,共表达了催化二硫键从头形成的巯基氧化酶(Erv1p)或/和促进二硫键正确折叠的二硫键异构酶(DsbC)。结果表明共表达DsbC可使C4ST-1融合蛋白的胞内可溶性表达水平显著提高,但C4ST-1和Erv1p共表达对胞内可溶性蛋白质的表达影响相对较小。C4ST-1和Erv1p或C4ST-1和DsbC共表达菌株的酶活分别为原始菌株的1.30和2.33倍,活力达到(12.32±0.76) U/L和(21.99±0.42) U/L。挑取同时共表达C4ST-1、Erv1p和DsbC的菌株进行摇瓶水平和3 L发酵罐放大培养,酶活分别达到(29.12±0.66) U/L和49.97 U/L。本研究为C4ST-1的大规模应用奠定了一定的基础。     

红色红曲菌组蛋白去乙酰化酶MrSir2的原核表达及活性分析

根据基因组序列信息,利用cDNA末端快速扩增技术(rapid-amplification of cDNA ends,RACE)得到了红色红曲菌中组蛋白去乙酰化酶mrsir2基因的完整cDNA序列,其编码序列(coding sequence,CDS)为1539 bp,编码512个氨基酸,含有一个SIR2蛋白保守结构域。根据大肠杆菌的密码子的偏好性对mrsir2序列进行优化,优化后的序列与p ET-28b载体连接后转入宿主菌E.coli BL21中进行IPTG诱导表达,并优化表达条件。实验结果表明:在16℃条件下用终浓度为0.25 mM的IPTG诱导培养16 h后,目的蛋白Mr Sir2可溶性表达效果良好。Ni2+柱亲和层析纯化后的Mr Sir2重组蛋白在SDS-PAGE上显示为一条约75 ku大小的条带,蛋白定量浓度达1.97 mg/mL,经Western blot鉴定为目的蛋白,测定酶活为78.5(OD/min/mg),780μM的二氢香豆素(dihydrocoumarin,DHC)对Mr Sir2蛋白的酶活抑制率为47%。Mr Sir2蛋白的可溶性表达为全面了解其酶学特征提供了材料,也为体外分析蛋白相互作用奠定了基础。