海洋红酵母Y2发酵产类胡萝卜素条件的研究

目的 对海洋红酵母Y2高产类胡萝卜素的发酵条件进行优化.方法 在摇瓶条件下,研究培养基成分和培养条件对海洋红酵母Y2生长和类胡萝卜素合成的影响,同时进行海洋红酵母Y2发酵过程的动态分析.结果 海洋红酵母Y2优化培养基组合为葡萄糖45 g/L,蔗糖15 g/L,酵母粉5 g/L,蛋白胨2.5 g/L,磷酸二氢钾1 g/L,磷酸二氢钠3 g/L,硫酸镁7.5 g/L,氯化钾3 g/L,氯化钠5 g/L.最适培养参数为:温度20℃,培养基初始pH为5,接种量为10％,250 mL摇瓶装液量为10～50 mL.类胡萝卜素的合成主要集中在对数生长期和稳定期.海洋红酵母Y2最适收获时间为72 h.种龄以36 h为宜.结论 利用优化培养基,在最适条件下培养海洋红酵母Y2,类胡萝卜素产量达到4.97 mg/L,比基础培养基提高了60.32％.     

Effectiveness of mouse minute virus inactivation by high temperature short time treatment technology: A statistical assessment

Viral contamination of mammalian cell cultures in GMP manufacturing facility represents a serious safety threat to biopharmaceutical industry. Such adverse events usually require facility shutdown for cleaning/decontamination, and thus result in significant loss of production and/or delay of product development. High temperature short time (HTST) treatment of culture media has been considered as an effective method to protect GMP facilities from viral contaminations. Log reduction factor (LRF) has been commonly used to measure the effectiveness of HTST treatment for viral inactivation. However, in order to prevent viral contaminations, HTST treatment must inactivate all infectious viruses (100%) in the medium batch since a single virus is sufficient to cause contamination. Therefore, LRF may not be the most appropriate indicator for measuring the effectiveness of HTST in preventing viral contaminations. We report here the use of the probability to achieve complete (100%) virus inactivation to assess the effectiveness of HTST treatment. By using mouse minute virus (MMV) as a model virus, we have demonstrated that the effectiveness of HTST treatment highly depends upon the level of viral contaminants in addition to treatment temperature and duration. We believe that the statistical method described in this report can provide more accurate information about the power and potential limitation of technologies such as HTST in our shared quest to mitigate the risk of viral contamination in manufacturing facilities.     

Inferring demographic structure with moccasin size data from the Promontory Caves,Utah

The moccasin assemblage Julian Steward recovered from the Promontory caves in 1930–31 provides a novel example in which material culture can be used to understand the structure of an AD thirteenth century population. Several studies shed light on the relationship between shoe size, foot size, and stature. We develop an anthropometric model for understanding the composition of the Promontory Cave population by using moccasin size as a proxy for foot size. We then predict the stature of the individual who would have worn a moccasin. Stature is closely related to age for children, subadults and adult males. Although there are predictable sex and age factors biasing moccasin discard practices, moccasin dimensions suggest a relatively large proportion of children and subadults occupied the Promontory caves. This bison and antelope hunting population appears to have thrived during its stay on Promontory Point. Am J Phys Anthropol 156:76–89, 2015 © 2014 Wiley Periodicals, Inc.     

Long‐term preservation of a collection of Rhizoctonia solani using cryogenic storage

Rhizoctonia solani is an important plant pathogen for a number of crops and maintaining an extensive collection of reference isolates is important in understanding relationships of this pathogen with multiple hosts. Current long‐term storage methods typically call for frequent transfer increasing the risk of changes in morphological, physiological or virulence characteristics. Cryopreservation using storage in liquid nitrogen (LN) was evaluated to examine the potential for storage of a R. solani culture collection containing 106 isolates (primarily from sugar beet). Cultures were stored on autoclaved barley grains in the vapour phase of LN. After 60 days, 5 years and 10 years in storage, all isolates were tested for viability by calculating the percentage of barley grains from which R. solani mycelia grew. Five years after initial storage, all isolates except one had no change in viability. After 10 years in storage, 67 of 106 isolates had no significant decrease in viability, 39 of 106 isolates had a significant decrease in viability but only 9 isolates had less than 10% growth, with 4 having no growth. A subset of isolates stored for 10 years were tested for pathogenicity on a susceptible (FC901) and resistant (FC703) sugar beet germplasm. All isolates tested maintained approximately the same level of virulence that they had prior to storage on both germplasms. This indicates that cryogenic methods are suitable for the preservation or storage of R. solani culture collections, although efficacy may vary with individual isolates.     

漆酶工程菌株的构建及其培养条件的研究

提取产漆酶白腐菌 (Fomelignosus)的总RNA ,利用RT_PCR的方法克隆到漆酶的cDNA ,并将其克隆到表达载体pPIC9k ,重组质粒经线性化、电激转化PichiapastorisGS115、通过底物显色反应筛选到漆酶生产工程菌株。其中 2 #工程菌株产酶活力较高。培养基、培养温度、诱导用甲醇浓度及培养基中铜离子浓度对该菌产酶活力均有一定的影响。在最适条件下 (用含 2 0 0 μmol LCu2 的BMMY培养基 ,于 2 0℃ ,2 5 0r min ;用 0 5 %甲醇诱导 )培养至第 6天时 ,该菌株产酶活力达到最高 (15 10U L)。     

Improvement of Fermentation Conditions for the Production of X-Prolyl-Dipeptidyl Aminopeptidase from Lactococcus Lactis

The quantitative effects of some fermentation conditions on the production of the enzyme X-prolyl-dipeptidyl aminopeptidase (PepXP)(EC 3.4.14.5) of Lactococcus lactis subsp. lactis and cremoris were studied. The PepXP activity was found both in the membrane and in the cytoplasm, suggesting the presence of multiple molecular forms. Both microorganisms showed higher PepXP activities when glucose (5 g/l) was used as the carbon source and the yeast extract in the culture medium was increased to 3.5 g/l. In these conditions, 226 mU/ml of PepXP activity were obtained with L. lactis subsp. lactis and 235 mU/ml with the subsp. cremoris after 6 h. The best fermentation temperature was in the 30–32 °C range. The enzyme activity remained stable even during the stationary phase.     

胎鼠脊髓源性神经干细胞分离培养与鉴定

目的:研究胎鼠的脊髓源性神经干细胞的分离培养方法并观察其增殖和分化能力。方法:利用显微操作技术分离获得胎鼠脊髓组织、无血清培养技术和酶消化法结合机械法传代培养神经干细胞、免疫细胞化学方法鉴定神经干细胞和分化情况。结果:建立了胎鼠脊髓源性神经干细胞的分离、培养和鉴定的方法,观察到了脊髓源性神经干细胞具有较强的增殖能力,在添加有5ng／mlEGF和5ng／mlbFGF的无血清培养液中可贴壁分化为神经元、少突细胞和星形胶质细胞。结论:在体外培养条件下分离培养的胎鼠脊髓源性神经干细胞具有干细胞的特性即较强的增殖能力和多向分化潜能。     

银杏根系甲醇溶提物对离体培养条件下丛枝菌根真菌生长发育的研究

银杏根系甲醇溶提物对离体培养条件下丛枝菌根真菌生长发育的试验结果表明,银杏根系甲醇溶提物对离体条件下丛枝菌根真菌(Glonus mosseae,Gigaspora margarita)生长发育有明显的促进作用,能显著提高孢子萌发率,增加菌丝生长长度。与对照相比,浓度为20％～100％的甲醇溶提物均可显著促进丛枝菌根真菌的生长发育,80％甲醇溶提物效果最明显,银杏根系甲醇溶提物中含丰富的黄酮类化合物,溶提物中黄酮类物质的含量与甲醇洗脱剂的浓度有关,当浓度为80％时,黄酮含量最高,对银杏根系甲醇溶提物中黄酮类化合物含量与丛枝菌根真菌生长发育相关性分析发现,溶提物中黄酮类化合物含量与离体条件下丛枝菌根真菌孢子的生长发育表现为极显著的正相关关系,溶提物中黄酮含量越高,其对丛枝菌根真菌孢子的生长发育促进作用越大。     

Strategies to improve the reliability of a theory: the experiment of bacterial invasion into cultured epithelial cells

An analysis is presented of published methods that have been used by experimenters to justify the reliability of the theory of invasion of microorganisms into cultured cells. The results show that, to demonstrate this invasion, many experimenters used two or more methods that were based on independent technical and theoretical principles, and by doing so improved the reliability of the theory. Subsequently I compare this strategy of ‘multiple derivability’ with other strategies, discussed in the literature in relation to the mesosome, a bacterial organelle that had been detected with the electron microsope, but which appeared later to be an artifact. I propose that different strategies have been applied in this problem, and multiple derivability may have been the decisive one. Finally I discuss the idea that multiple derivability may help to anchor theories in a larger network of theories.