Xp11.2区域YAC OAT1中表达顺序筛选

筛选 X染色体 Xp11.2区域的酵母人工染色体 ( YAC)克隆 OAT1所包含的表达序列 ,为该区域疾病相关基因的克隆探索方法。以 YAC克隆 OAT1为探针 ,与胎脑 c DNA文库的噬菌体原位裂解膜进行两轮杂交。所得的阳性克隆经过杂交鉴定剔除假阳性克隆和重复序列 ,并进行测序分析。获得了一个 YAC OAT1编码的表达顺序克隆。该克隆与鸟氨酸转氨酶 ( Ornithine Amino-transferase,OAT)基因同源     

Bioassay‐directed fractionation for analyzing estrogens in surface waters of the German Baltic Sea

A reversed‐phase fractionation method with subsequent biological and chemical analysis has been developed to estimate the contributions of the most potent estrogens to observed estrogenic effect potentials. Surface water samples were taken in the German Baltic Sea (Inner Wismar Bay and Darss Peninsula, sampling campaign July 2003) and were separated into seven individual fractions. Three fractions showed significant estrogenic activities and clear dose‐dependant responses were obtained in the yeast estrogen screen (YES). In the 2nd fractions liquid chromatographic‐electrospray‐tandem mass spectrometric (LC‐ESI‐MS‐MS) analyses showed the presence of bisphenol A (Inner Wismar Bay: 4.8 ng L^–1 and 6 ng L^–1; Darss Peninsula: 0.91 ng L^–1 and 1.7 ng L^–1) and ethinylestradiol (Inner Wismar Bay: 2.0 ng L^–1 and 6.0 ng L^–1; Darss Peninsula: < MDL and 1.7 ng L^–1), whereas estrogenic activities in the YES were only around 10% of the positive control E2. Although not identified prior in the total extract the natural hormones estradiol (Inner Wismar Bay: 0.13 ng L^–1 and 0.19 ng L^–1; Darss Peninsula: 0.12 ng L^–1 and 0.16 ng L^–1) and estriol (Inner Wismar Bay: < MDL and 0.33 ng L^–1; Darss Peninsula: < MDL) could be detected in the 3rd fractions, where high estrogenic potentials could be observed. The 4th fractions showed high responses as well and estrone were herein quantified with concentrations of 0.16 ng L^–1 and 0.18 ng L^–1 (Darss Peninsula) up to 0.37 ng L^–1 (Inner Wismar Bay). Measured and calculated estradiol equivalents for individual fractions correlated very well (R² = 0.78), when disregarding results of the 2nd fraction, where high deviations occurred.     

转鲑鱼生长激素基因酵母菌发酵特性的研究

张培军、Ball等研究表明,外源性生长激素通过注射或浸泡的方法可加快鱼的生长[1]。根据Mclean1987年、Shunsuke等1993年研究发现鱼类后肠具有吸收部分生长激素进入血液循环的特点,本研究室将鲑鱼生长激素基因克隆进无毒无害的酵母菌中,利用可控制的诱导条件,培养出含鲑鱼生长激素的酵母菌,直接掺入饵料,使生长激素通过消化道吸收,以促进鱼的生长。Jacqueline等1989年指出,由于携带外源基因的酵母菌与普通酵母菌的培养要求不同,既要促使菌体快速生长又要保证外源基因的表达,本研究的目的在于选择适宜的培养条件,实现转基因酵母菌的高密度培…     

三株海洋酵母的生化营养成分分析

海洋酵母是一种重要的水产生物饵料.实验对3株海洋酵母菌德巴利酵母Debaryomyces sp.、克鲁维酵母Kluyveromyces sp. 和虫道酵母Ambrosiozyma sp.的基础营养成分进行了研究和分析.3株海洋酵母菌的粗蛋白含量分别为56.87%、46.75%、51.80%;脂肪分别为4.67%、6.35%、6.35%;碳水化合物分别为31.28%、41.55%、35.96%;灰分分别为7.18%、5.35%、6.37%.3株海洋酵母菌具有较高的蛋白含量水平以及优良的氨基酸和脂肪酸组成,表明该3株海洋酵母菌具有用作水产饵料生物的营养价值.但是,3株海洋酵母菌均缺乏多不饱和脂肪酸EPA和DHA,用其作为水产饲料源料,需要在配合饲料中添加多不饱和脂肪酸或与富含多不饱和脂肪酸的藻类一起使用.     

Amylase Production by the Marine Yeast Aureobasidium pullulans N13d

1 Introduction Amylases are enzymes which hydrolyze starch molecules to give diverse products including dextrin and pro-gressively smaller polymers composed of glucose units(Windish et al., 1965; Pandey et al., 2000; Chi et al.2001). Amylase is a kind of very important enzyme andconstitutes a class of industrial enzymes sharing approximately 25% of the enzyme market (Sindhu et al.1997; Rao et al., 1998). Amylases are universally distributed throughout the animal, plant and microbial kingdoms…     

Amylase production by the marine yeast Aureobasidium pullulans N13d

The marine yeast strain N13d, producing an extracellular amylase, was isolated from the deep sea sediments of the Pacific Ocean. This strain was identified to be Aureobasidium pullulans by 18S rRNA gene sequence analysis and routine yeast identification methods. The optimal sea water medium for amylase production by this yeast strain was 1.0% peptone and 1.0% soluble starch with pH 4.0. The optimal conditions for amylase production by this yeast strain were with temperature 28 °C, aeration rate 6 Lmin⁻¹ and agitation speed 250 rmin⁻¹. Under these conditions, 58.5 units of amylase activity per mg protein were produced within 56 h of fermentation.     

可口革囊星虫(Phascolosoma esculenta)酵母双杂交文库和铁结合蛋白真核表达载体的构建

构建了可口革囊星虫(Phascolosoma esculenta)的酵母双杂文库, 总克隆数为6.09×106CFU, 重组率100%, 平均插入片段长度>1kb。随机挑取200个克隆子测序, 结果1个空载, 67 个无法对应的基因类型, 已知的132个基因中线粒体蛋白占29.54%, 蚯蚓血红蛋白15.9%, 核糖体蛋白10.6%, 铁结合蛋白8.3%, 肌动蛋白6.1%, 其它基因29.56%。这些基因分别为: 5-氨基乙酰丙酸合成酶、类博莱霉素水解酶、纤溶蛋白、谷氨酰胺合成酶、热休克蛋白、非特性类碱性磷酸酶、脂肪酶、金属蛋白、线粒体、硫氧还蛋白、蛋白磷酸酶、肌钙蛋白C、泛素。根据已获得的可口革囊星虫铁结合蛋白基因, 以EcoRⅠ和KpnⅠ为双酶切位点设计引物, 构建了可口革囊星虫铁结合蛋白真核表达载体pPink-HC-fer, 经PCR 及测序检验, 序列完全正确, 然后电转化至酵母细胞中诱导表达, 经SDS-PAGE检验其融合蛋白分子量大小约为23kDa。     

重组鲑鱼降钙素结肠黏附缓释微球的制备

为考察重组(酵母)鲑鱼降钙素结肠黏附缓释微球的制备工艺和最优处方,并对释药特性进行探讨。以阿拉伯胶、β-环糊精、可溶性淀粉、明胶为壁材,HPMC和卡波姆为生物黏附剂,运用正交试验确定喷雾干燥法制备微球的最佳工艺条件和处方。结果表明,微球的最佳制备工艺和处方为β-环糊精与阿拉伯胶以15∶1、HPMC与CP以4∶1配比为理想壁材,喷雾干燥进风温度160℃、进料速度10mL·min-1、雾化压力0.5MPa、壁材浓度15%、芯材比1∶3为最佳组合,所得微球能达到缓释12h的试验设计要求。本法工艺稳定、可行,所得鲑鱼降钙素结肠黏附缓释微球具有良好的缓释效果。     

Differences in population growth of rotifers and cladocerans raised on algal diets supplemented with yeast

We studied the population growth of two rotifer (Brachionus calyciflorus and Brachionus rubens) and two cladoceran (Ceriodaphnia dubia and Moina macrocopa) species fed three diets (Chlorella vulgaris (Ch), Scenedesmus acutus (Sc) and yeast (Saccharomyces cerevisiae) (Y) in seven combinations (alone or mixed in equal proportions (on dry weight basis): Ch+Sc, Ch+Y, Sc+Y and Ch+Sc+Y). In general, the cladocerans were more adversely affected than the rotifers on the diet of yeast alone. The population growth curves of B. calyciflorus and B. rubens revealed that algal diets were superior to yeast. Regardless of diet, B. rubens had a longer lag phase and delayed peak density compared to B. calyciflorus. Both M. macrocopa and C. dubia had higher peak abundances when fed mixed algae than on either alga offered separately. B. rubens, C. dubia and M. macrocopa reached significantly lower maximal densities on diets containing yeast, S. acutus or both. When the data on the eggs/female (egg ratio) were plotted as function of population density, we found an inverse relation, which was curvilinear for B. calyciflorus and linear for B. rubens. In general, we found that yeast could effectively supplement algal diets in all the test species, thereby reducing costs in large scale production of zooplankton.     

云南高原湖泊抚仙湖酵母菌空间分布及其与环境因子的关系

开展高原湖泊酵母菌多样性研究,能够为湖泊生态系统的保护提供理论依据,并为其中特殊酵母菌资源的开发及利用奠定基础.结合经典分类法及26S rDNA D1/D2区域序列分析,对分离自云南抚仙湖湖水中的553株酵母菌进行系统分类,运用SPSS 19.0软件比较不同区域酵母菌多样性,并采用多元统计方法定量分析酵母菌空间分布特征及其与理化因子之间的关系.结果显示:抚仙湖水体中分布22属52种和1个潜在新分类单元的酵母菌.理化因子差异性分析表明,北沿岸区总有机碳浓度明显高于南沿岸区.Pearson分析则表明,抚仙湖湖水总有机碳浓度与酵母菌丰度呈显著正相关.另外,酵母菌-环境冗余分析显示,抚仙湖酵母菌种群结构与总有机碳浓度存在明显相关.研究显示云南抚仙湖酵母菌资源比较丰富,人类活动对其中酵母菌空间分布具有一定影响.