引入非天然氨基酸胶原蛋白表达及交联成键的优化

微生物重组表达胶原蛋白来源清洁,同时具有序列设计灵活和高产量高纯度等优点,作为生物材料在组织工程等领域具有广泛的应用前景。然而如何促进重组胶原分子交联,使其形成更加稳定的空间结构是设计重组胶原纳米材料需要克服的难点。文中通过双质粒系统将非天然氨基酸O-(2-溴乙基)-酪氨酸引入细菌胶原蛋白序列中,并对其发酵条件进行优化,结果表明在25 ℃下,以终浓度为0.5 mmol/L的IPTG和0.06%的阿拉伯糖诱导24 h可以获得高纯度含非天然氨基酸的胶原蛋白。将含非天然氨基酸的胶原蛋白与含半胱氨酸的胶原蛋白在pH为9.0的NH4HCO3缓冲液中进行交联,形成了最大分子粒径可达1 μm的聚集体,为重组胶原蛋白生物材料的设计提供了新思路。     

Recent developments in membrane-protein structural genomics

Recent work has identified the topology of almost all the inner membrane proteins in Escherichia coli, and advances in nuclear magnetic resonance spectroscopy now allow the determination of α-helical membrane protein structures at high resolution. Together these developments will help overcome the current limitations of high-throughput determination of membrane protein structures.     

海南省香蕉枯萎病病原菌的分离鉴定及1号、4号小种的生物学特性

香蕉枯萎病是目前国内和世界很多地区香蕉种植区的一种毁灭性病害,目前尚未找到有效的防治方法.本文采集了海南省不同地区香蕉枯萎病的病原菌,通过经典病理学与分子生物学的方法进行了鉴定,此外还优化了快速检测的方法.结果表明,通过接种巴西香蕉和粉蕉进行致病性检测以及分离菌rDNA-ITS测序结果,明确鉴定出供试的香蕉分离菌株为4号生理小种,粉蕉分离菌株为1号生理小种.为了对1号、4号生理小种有更多的了解,本文还对1号、4号小种菌株生物学特性进行了测定,确定了影响菌丝生长的最佳条件.结果显示,pH 5时最适合1号、4号生理小种的菌丝生长,适合菌丝生长的温度为19～28℃之间,温度达到37℃时菌丝不能生长,碳源对菌丝的生长影响不大.这些方法和数据可以有利于田间香蕉枯萎病病害的检测和最终确定,生物学特性的测定可以为耕作技术的改进和农药的研发提供参考依据.     

不同冷季型牧草品种的产草量及品质比较分析

采用随机区组试验设计,对引进的8个冷季型牧草品种进行产草量和营养成分的比较分析.结果表明:4次刈割的总鲜重和总干重在品种间差异极显著,且以蓝天堂黑麦草(7号)最高.品种间的粗蛋白含量差异显著,其中第1次刈割以奇可利(3号)最高,蓝天堂黑麦草(7号)居第2位;第2次仍以奇可利(3号)最高,特高黑麦草(1号)和蓝天堂黑麦草(7号)居其次;第3次以紫花苜蓿(8号)最高,其次是多年生黑麦草(4号);第4次以奇可利(3号)最高,多年生黑麦草(4号)居第2位;粗纤维、粗脂肪和粗灰分在品种间的差别较小.综合产草量和品质两方面考虑,蓝天堂黑麦草(7号)可作为本地首选推广的冷季型牧草品种,奇可利(3号)和多年生黑麦草(4号)可作为备选推广的牧草品种.     

Characterization of the carboxysomal carbonic anhydrase CsoSCA from Halothiobacillus neapolitanus

In cyanobacteria and many chemolithotrophic bacteria, the CO(2)-fixing enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) is sequestered into polyhedral protein bodies called carboxysomes. The carboxysome is believed to function as a microcompartment that enhances the catalytic efficacy of RubisCO by providing the enzyme with its substrate, CO(2), through the action of the shell protein CsoSCA, which is a novel carbonic anhydrase. In the work reported here, the biochemical properties of purified, recombinant CsoSCA were studied, and the catalytic characteristics of the carbonic anhydrase for the CO(2) hydration and bicarbonate dehydration reactions were compared with those of intact and ruptured carboxysomes. The low apparent catalytic rates measured for CsoSCA in intact carboxysomes suggest that the protein shell acts as a barrier for the CO(2) that has been produced by CsoSCA through directional dehydration of cytoplasmic bicarbonate. This CO(2) trap provides the sequestered RubisCO with ample substrate for efficient fixation and constitutes a means by which microcompartmentalization enhances the catalytic efficiency of this enzyme.     

中国绿杯菌属研究(英文)

对采自中国12个省的绿杯菌属Chlorociboria标本进行了分类研究,鉴定出3个种,分别为小孢绿杯菌C.aeruginascens、绿杯菌C.aeruginosa和波托绿杯菌C.poutoensis。其中波托绿杯菌为中国新记录种,对该种的形态学特征进行了描述和图示,并对其分类地位提供了DNA序列分析的佐证。     

云丘山地区灌木群落优势种数量关系

以68个灌木样方的重要值矩阵和2×2列联表为基础,结合实地调查,应用方差比率法、χ2检验、Pearson相关系数及Spearman秩相关系数法,对云丘山地区灌木群落的20个优势种、190个种对的关联性进行了定量研究。方差比率法结果表明,20个优势种总体间关联为显著正相关,云丘山地区的灌木群落处于较稳定的状态;χ2检验、Pearson相关系数、Spearman秩相个关系数结果表明,大部分种对的种间关联未达到显著程度,种间联结较为松散,群落结构及其种类组成将逐渐趋于完善和稳定;根据这20个优势种对环境的适应方式和主导生态因素辅助以PCA排序,可将它们划分为两种生态种组。以上结果揭示了该地区主要优势物种与环境的相互关系。     

酱香型白酒酿造酒醅中酵母菌多样性研究

为解析酱香型白酒酿造酒醅中酵母菌的菌群结构,获取酒醅中的主要酵母菌,采用高通量测序法分析酱香型白酒酒醅中酵母菌多样性及主要功能菌群,同时采用可培养分离方法获取酒醅中酵母菌活性菌株。从酱香型白酒下沙至五轮次酒醅中共检出59个属、129个种的酵母菌,分离得到酵母菌活性菌株41种,检测到的酵母菌种类与获得的酵母菌活菌在各香型白酒中最多。不同时期酒醅中的酵母菌种类和数量差异明显,其中下沙、造沙轮次以Pichia kudriavzevii为绝对优势酵母菌;一至五轮次随着轮次的递增,酒醅中优势酵母菌的种类增多,其中主要的优势酵母菌有Pichia kudriavzevii、Pichia manshurica、Zygosaccharomyces bailii、Saccharomyces cerevisiae、Candida apicola。酱香型白酒酒醅中蕴藏着极其丰富的酵母菌资源,对酵母菌菌群结构的解析有助于科学地认识酱香型白酒酿造过程中产酒与风味代谢机理,为发酵过程的调控提供一定依据。     

新生大鼠下丘脑神经元L-Ca~(2 )通道电流活动特征

目的 :研究新生大鼠下丘脑神经元L Ca2 通道单通道特性 ;Ca2 通道激动剂BayK 86 44对Ca2 通道单通道特性的影响。方法 :采用神经元急性分离技术 ;用膜片钳细胞贴附式记录方式进行研究。结果 :大鼠下丘脑神经元L Ca2 通道是一种电导相对较大的Ca2 通道 ,其电导为 (2 9.5± 3.1)pS ,平均开放时间 (τ0 )为 0 .2 8ms,平均关闭时间的短关闭时间常数 (τc1)为 2 .91ms,长关闭时间常数 (τc2 )为 5 3.2 2ms。此通道几乎不存在时间依赖性失活。BayK86 44显著增加通道的开放概率 ,通道平均开放时间增加为 1.6 1ms。结论 :下丘脑神经元存在L Ca2 通道 ,该通道具有明显电压依赖性 ,而无显著的时间依赖性。通道特征与文献报道的其它神经元上L Ca2 通道相似 ,也有明显不同 ,显示下丘脑神经元L Ca2 钙通道的独特性     

A green and effective method for the determination of metalaxyl enantiomers in tobacco and soil by supercritical fluid chromatography–tandem mass spectrometry

We reported a new methodology for the stereoselective determination of metalaxyl enantiomers in tobacco and soil. The QuEChERS (quick, easy, cheap, effective, rugged, and safe) method was used for the extraction and clean-up of the tobacco and soil samples. Separation of the metalaxyl enantiomers was performed on an ACQUITY UPC2 Trefoil CEL1 chiral column coupled with supercritical fluid chromatography with tandem mass spectrometry (SFC-MS/MS), and the run time was only 5 minutes. Under the optimized conditions, the recoveries for the enantiomers were between 78.2% and 93.3% with intraday relative standard deviations (RSDs) ranging from 1.1% to 5.4%. The limit of detection (LOD) for the enantiomers in tobacco and soil varied from 0.005 to 0.007 mg/kg, and the limit of quantitation (LOQ) ranged from 0.017 to 0.020 mg/kg. In this method, only a small amount of methanol was consumed to obtain a rapid stereoselective separation. This proposed method showed good accuracy and precision and might be suitable for fast enantioselective determination of metalaxyl in food and environmental samples. The developed method was further validated by application to the analysis of authentic samples.