Isolation and characterization of peptide antibiotics LI-F04 and polymyxin B6 produced by Paenibacillus polymyxa strain JSa-9

Paenibacillus polymyxa JSa-9 had been found to produce five cyclic LI-F type antibiotics which were released into culture medium in accordance with our previous report. In this study, another three kinds of antagonistic compounds were extracted from P. polymyxa JSa-9 cell pellets and (or) spores by methanol. Using high performance liquid chromatography (HPLC) method, two antagonistic fractions were separated and collected from the methanol extract. One showed inhibition against Escherichia coli and Staphylococcus aureus, while the other was active against Aspergillus niger and S. aureus. By means of electrospray ionization mass spectroscopy (ESI-MS), infrared spectroscopy (IR), and amino acid analysis, two kinds of compounds from fraction B with molecular masses of 901 and 915 Da were characterized as the linear lipopeptide analogs of antibiotics LI-F04a and LI-F04b, respectively. Another antimicrobial substance from fraction A could be attributed to polymyxin B₆.     

糖皮质激素与脂肪代谢和胰岛素抵抗

内源性和外源性糖皮质激素过多会导致胰岛素抵抗和发展为糖尿病.糖皮质激素增加循环中葡萄糖、胰岛素和游离脂肪酸(FFA)浓度.循环中FFA增高与肥胖、胰岛素抵抗和2型糖尿病密切相关,其主要来源于脂肪细胞内甘油三酯水解.糖皮质激素刺激脂肪分解、增加FFA构成了激素导致胰岛素抵抗的重要机制之一.     

大肠杆菌热休克或冷休克启动子调控的基因表达载体

基因重组技术已经成为获得各种酶和生物活性蛋白的主要手段。虽然很多基因已在大肠杆菌中得到高效表达,但是当人们认定某种蛋白对科学研究或生产应用极为重要时,却常常因为其基因表达水平很低或产生包涵体而感到束手无策。表达载体pHsh和pEXC通过激活热休克或冷休克转录调控机制提高分子伴侣的表达水平,从而降低目标蛋白的细胞毒性并减少包涵体形成。应用于生物合成、分子修饰或生物降解的高温酶可以通过pHsh系统表达获得高产,而科研和诊疗所需要的来源于动植物和常温微生物的基因可以通过pEXC系统获得高效表达。这些新载体的发展为重组蛋白的小规模制备和大规模生产提供了新策略和有效途径。     

SRAP、ISSR技术的优化及在甘蓝类植物种子鉴别中的应用

将SRAP与ISSR 2种分子标记技术应用于8种甘蓝类植物（Brassica oleracea L.）的种子鉴别中。先以甘蓝（Brassica oleracea var. capitata）基因组DNA为模板,通过对SRAP、ISSR反应体系中各影响因素的逐一筛选,优化了甘蓝类植物SRAP、ISSR反应体系。进而采用30个SRAP引物组合和15个ISSR引物对白甘蓝、皱叶甘蓝、红甘蓝、羽衣甘蓝、花椰菜、青花菜、抱子甘蓝、球茎甘蓝的基因组DNA进行了PCR扩增,结果表明：M3E5与M4E5两个SRAP引物组合可以在8种甘蓝类植物之间显示较高的多态性;844和888两个ISSR引物也可在8种甘蓝类植物之间产生很好的多态,特别是844引物单独应用即可区分所有材料。     

黄河三角洲植物群落分布格局及其影响因素

利用双向指示种分析和去趋势典范对应分析的方法,对典型河口滨海湿地黄河三角洲植物群落类型和结构进行了分析,并探讨了植物群落分布格局及其影响因素。结果表明：134个样方可划分为7个群系,即刺槐群落（Form. Robinia pseucdoacacia）、荻群落（Form. Miscanthus saccharifleus）、翅碱蓬群落（Form. Suaeda heteroptera）、獐毛群落（Form. Aeluropus sinensis）、芦苇群落（Form. Phragmites communis）、柽柳群落（Form. Tamarix chinensis）和白茅群落（Form. Imperata cylindrica）。群落类型的划分主要反映了微地貌所决定的地下水埋深及土壤盐分含量的变化。在影响植物群落分布格局的因素中,环境因子解释的部分占45.2%,空间因子解释的部分占11.8%,空间因子和环境因子交互作用解释的部分占2.23%,空间因子和环境因子未能解释的部分占40.8%,这部分主要反映了群落自身互作及人类活动对群落的影响。     

Lymphocyte-specific Ca2+-binding protein LSP1 is associated with the cytoplasmic face of the plasma membrane.

The gene for LSP1 is a lymphocyte-specific gene previously isolated by us using a subtractive hybridization technique. LSP1 mRNA is found in normal and transformed B lymphocytes and in normal T lymphocytes but not in transformed T lymphocytes. To study the expression of the mouse LSP1 protein, we prepared a polyclonal antiserum specific for the LSP1 protein. Here we report that the gene for LSP1 was expressed in transformed B-lymphoma cell lines and in normal mouse thymocytes as a protein doublet with apparent molecular masses of 52 and 50.5 kilodaltons when analyzed on a sodium dodecyl sulfate-10% polyacrylamide gel. BW5147 cells transfected with an LSP1 cDNA clone expressed only the 52-kilodalton protein. No LSP1 protein was expressed in nine T-lymphoma cell lines tested. Immunofluorescence studies of intact and permeabilized cells and subcellular fractionation experiments showed that the LSP1 protein was associated with the cytoplasmic side of the plasma membrane in transformed B-lymphoma cell lines and in normal thymocytes. Using a simple filter-binding assay, we showed that recombinant LSP1 protein was Ca2+ binding, as predicted on the basis of its deduced amino acid sequence. On the basis of the particular expression pattern, the subcellular localization, and the Ca2+-binding property of the LSP1 protein, we hypothesize that the LSP1 protein is a lymphocyte-specific component of a signal transduction pathway involved in the regulation of lymphocyte growth.     

大鼠侧脑室慢性注射CRF致抑郁样变化研究

为了探讨CRF在抑郁症发生发展过程中的作用．对正常大鼠侧脑室慢性注射CRF21天并与慢性非预见性应激刺激21天建立的抑郁症模型大鼠进行比较。运用旷场行为实验（open-field）观察大鼠主动性活动能力．用Morris water Maze法．以训练期的逃避潜伏期为指标检测大鼠空间学习记忆能力。采用HPLC—UV法测定血清皮质醇含量,RT—PCR法检测CRF及其受体mRNA的表达。结果显示：慢性应激21天建立的模型大鼠主动性活动和学习记忆能力均明显下降．血清皮质醇含量显著升高,CRF及其受体R1 mRNA的表达增加。大鼠侧脑室慢性注射CRF21天后．其体重增量、主动性活动和学习记忆能力与慢性应激模型大鼠一样均明显降低。这些工作证明了CRF在抑郁症的发生发展过程中发挥了至关重要的作用．慢性应激导致机体CRF分泌持续增加可能是抑郁症发病的主要原因。     

人血清白蛋白-睫状神经营养因子突变体融合蛋白基因在毕赤酵母中的表达及表达产物的纯化和活性鉴定

为了延长人睫状神经营养因子突变体的体内半衰期 ,将人血清白蛋白 (HAS)的C 末端和人睫状神经营养因子突变体AX15 (R13K)的N 末端通过一个 11个氨基酸的连接肽融合在一起 ,构建了融合蛋白HAS-AX15 (R13K)。HAS-AX15 (R13K)融合蛋白基因在巴斯德毕赤酵母中进行表达后通过阳离子交换层析、反向层析和凝胶过滤对表达产物进行了分离纯化。体外TF 1细胞存活实验表明与HAS融合并未影响AX15 (R13K)的生物学活性。体内动物实验表明HAS-AX15 (R13K)融合蛋白的疗效比AX15 (R13K)更为持久 :每 3天注射一次 4 8mg/kg的HAS-AX15 (R13K)融合蛋白的治疗效果优于每天注射一次 1 6mg/kg的AX15 (R13K)的治疗效果。HAS-AX15(R13K)融合蛋白不但可以减少用药次数 ,提高病人的顺应性 ,而且还可以减少用药量和血药浓度的波动 ,从而降低副反应 ,在临床应用上具有一定的优势。     

Engineering flavonoid glycosyltransferases for enhanced catalytic efficiency and extended sugar-donor selectivity

Flavonoids are predominantly found as glycosides in plants. The glycosylation of flavonoids is mediated by uridine diphosphate-dependent glycosyltransferases (UGT). UGTs attach various sugars, including arabinose, glucose, galactose, xylose, and glucuronic acid, to flavonoid aglycones. Two UGTs isolated from Arabidopsis thaliana, AtUGT78D2 and AtUGT78D3, showed 89 % amino acid sequence similarity (75 % amino acid sequence identity) and both attached a sugar to the 3-hydroxyl group of flavonols using a UDP-sugar. The two enzymes used UDP-glucose and UDP-arabinose, respectively, and AtUGT78D2 was approximately 90-fold more efficient than AtUGT78D3 when judged by the k _cat/K _m value. Domain exchanges between AtUGT78D2 and AtUGT78D3 were carried out to find UGTs with better catalytic efficiency for UDP-arabinose and exhibiting dual sugar selectivity. Among 19 fusion proteins examined, three showed dual sugar selectivity, and one fusion protein had better catalytic efficiency for UDP-arabinose compared with AtUGT78D3. Using molecular modeling, the changes in enzymatic properties in the chimeric proteins were elucidated. To the best of our knowledge, this is the first report on the construction of fusion proteins with expanded sugar-donor range and enhanced catalytic efficiencies for sugar donors.