一株蟾毒灵转化菌株的筛选鉴定与转化条件优化

【背景】微生物转化是对天然产物进行结构修饰的重要手段,具有催化反应快、选择性强、反应条件易控制和污染小等特点。许多微生物能够对蟾毒配基类化合物进行生物转化,在不同位置进行结构修饰,并且产生毒性减弱的活性衍生物。【目的】筛选蟾毒灵生物转化菌株,以期发现活性更好、毒性降低的化合物。【方法】利用蟾毒灵为底物筛选其生物转化菌株,通过HPLC (High performance liquid chromatography)/LC-MS (Liquid chromatograph-mass spectrometer)鉴定转化产物;对可生物转化蟾毒灵的菌株进行菌落形态观察和分子生物学鉴定,并优化发酵条件提高转化率,同时测试该菌株对其他甾体化合物的转化作用。【结果】筛选获得一株蟾毒灵转化菌,经形态学与ITS (Internal transcribed spacer)分析鉴定为Naganishia属菌,转化产物为3-去氢蟾毒灵。该菌株生物转化的培养基最适底物浓度为8 mg/L,最适初始pH值为6.5,最佳接种量为3%,最佳转化时间为96 h,最终转化率为48.3%。该菌株可将雌酮E1转化为雌二醇E2,也可将雌二醇E2逆向转化为E1。【结论】首次发现Naganishia属菌株对甾体类化合物具有转化活性,其产生的弱细胞毒性的转化产物3-去氢蟾毒灵有望成为高效、安全的强心药物。微生物转化法选择性高、反应条件温和、操作简单,为大量制备活性化合物3-去氢蟾毒灵提供了一条简便易行的道路,也为更多甾体化合物结构修饰提供了可能。     

东方蝾螈原肠形成期瓶状细胞的结构及其对胚胎发育的影响

以１００只东方蝾螈早期胚胎为材料,用ＳＥＭ和ＴＥＭ对瓶状细胞形态结构和除去瓶状细胞的胚胎进行观察,去除瓶状细胞后,胚胎的原肠形成不会中断,外包和内卷仍能正常进行,并可以发育成幼螈,但有部分胚胎原肠前端缺失,长期培养的蝾螈胚对针刺激有反应,幼虫阶段中自由活动,我们的结论是：１）东方蝾螈原肠胚的瓶状细胞除了原肠前端存在外,还有部分瓶状细胞与原肠长轴垂直,从力学角度推断瓶状细胞在原肠形成过程中不起主要作     

北京市怀沙-怀九河市级水生野生动物保护区鱼类物种多样性及其资源保护

怀沙河和怀九河是两条自然汇入北京怀柔水库的河流,被规划为北京市第一个市级水生野生动物保护区。2004年3月~2005年2月,作者对该保护区内野生鱼类资源进行了本底调查。调查区域内野生淡水鱼类共计24种(不计引入种),隶属于6目11科24属,以鲤形目鲤科鱼类为主,多为山区溪流小型鱼类。鱼类物种多样性在空间分布上有所不同,两河下游近水库河段物种多样性最高,这可能与两个不同生态环境交界处的边缘效应有关;其多样性在时间上春夏季高于秋冬季。结合区域内的生境特点,建议加强对核心区的保护力度,重新规划缓冲区和过渡区;在保护自然环境的同时,兼顾当地居民的经济利益,达到人与资源的和谐发展。     

多媒体在微生物教学中的应用

多媒体计算机技术教学是一种新型的教学方式之一。多媒体教学克服了传统的微生物教学可视化差、形象不够直观等缺点。但全课堂多媒体教学则会扼杀教师教学中的即兴创造,成为典型的电脑“满堂灌”。所以多媒体课件的制作应符合教学规律,片面强调声形兼备,会导致许多环节与课堂脱节。教学中应目的明确、化难为易,探索其固有教学规律,努力做到教学相长。     

太白红杉群落物种多样性与环境因子的关系

定量研究了太白红杉群落物种多样性与环境因子的关系,结果表明:(1)海拔与物种丰富度和种间相遇机率呈极显著的负相关;与Shannon指数、Simpson指数和Pielou指数表现为"中间高度膨胀"的规律,即中等海拔高度上多样性高而低海拔和高海拔多样性较低。(2)岩石裸露度与物种丰富度呈显著的负相关。(3)土壤含水量与物种丰富度呈极显著的正相关;与种间相遇机率呈先降后升的趋势。(4)环境因子与物种多样性逐步回归结果是:海拔和岩石裸露度与物种丰富度关系密切,其回归方程为:S=90.62-0.02E-14.14B(r=0.769,P<0.05)。     

Isolation and analyses of genes preferentially expressed during early cotton fiber development by subtractive PCR and cDNA array

Cotton fibers are differentiated epidermal cells originating from the outer integuments of the ovule. To identify genes involved in cotton fiber elongation, we performed subtractive PCR using cDNA prepared from 10 days post anthesis (d.p.a.) wild-type cotton fiber as tester and cDNA from a fuzzless-lintless (fl) mutant as driver. We recovered 280 independent cDNA fragments including most of the previously published cotton fiber-related genes. cDNA macroarrays showed that 172 genes were significantly up-regulated in elongating cotton fibers as confirmed by in situ hybridization in representative cases. Twenty-nine cDNAs, including a putative vacuolar (H⁺)-ATPase catalytic subunit, a kinesin-like calmodulin binding protein, several arabinogalactan proteins and key enzymes involved in long chain fatty acid biosynthesis, accumulated to greater than 50-fold in 10 d.p.a. fiber cells when compared to that in 0 d.p.a. ovules. Various upstream pathways, such as auxin signal transduction, the MAPK pathway and profilin- and expansin-induced cell wall loosening, were also activated during the fast fiber elongation period. This report constitutes the first systematic analysis of genes involved in cotton fiber development. Our results suggest that a concerted mechanism involving multiple cellular pathways is responsible for cotton fiber elongation.     

一株新城疫病毒新强毒株(NL)的分离鉴定及F基因序列测定

１９９６年５月以来,１我国各地鸡群暴发鸡新城疫,其发病特征是常规疫苗免疫无效。为研究此病毒的特征,对从华东某地分离的新城疫病毒毒株（ＮＬ株）进行了生物学研究。利用ＲＴ－ＰＣＲ技术扩增出ＮＬ株融合蛋白基因（Ｆ基因）全序列,并测序。结果表明,ＮＬ株平均致鸡胚死亡时间最短的,其尿囊液中病毒滴度可达２＾９,表明病毒繁殖速度快。Ｆ基因测序结果表明,ＮＬ株为一强毒株,与现今国外已发表的ＮＤＶ株Ｆ基因同源性在８     

鲈鱼生长激素的分离及其生物活性鉴定

运用葡聚糖凝胶Ｇ－１００过滤和反相高儿液相色谱纯化两步法,首镒从鲈鱼脑垂体中分离出鲈鱼生长激素,通过ＳＤＳ－聚丙烯凝胶电泳测得鲈鱼非还原性的和还原性的生长激素分子量分别为１９．２和２０．７ｋＤ;等电聚焦证实鲈鱼生长激素等电点为７．１５。Ｗｅｓｔｅｒｎ免疫印迹反应证实,鲈鱼生长激素具有与大麻哈鱼生长激素抗体发生特异性免疫交叉反应的特性,而与大麻哈鱼催乳素和生长催乳素抗体无免疫交叉反应。     

Immunolcocalization of actin in intact and DNA—and histone—depleted nuclei and chromosomes of allium cepa

The presence of actin in eukaryotic nuclei and chromosomes,and especially in higher plant nuclei and chromosomes,has not been well established.We detected actin in meristematic cells of Allium cepa with indirect immunofluorescence technique and observed bright fluorescence in the intact nuclei and chromosomes,indicating that actin is present in the nuclei and chromosomes of the higher plant.We labeld sections of the meristematic cells of A.cepa with immunogold technique,gold parti cles were concentrated in condensed chromatin and nucleoli,confirming the results of the immunofluoresence observations.We traeated the nuclei and chromosomes of A.cepa with DNase I and 2M NaCl and obtained DNA-and histone-depleted nuclei and chromosomes.Indirect immunofluorescence tests showed that the DNA-and histonedepleted nuclei and chromosomes reacted positively with the anti-actin antibodies.These results demonstrate that the anti-actin antibodies.These results demonstrate that actin exists not only in intact nuclei and chromosomes,but also in DNA-and histone-depleted nuclei and chrmosomes of the plant.In addition,our immuno-fluorescence tests indicate that tropomyosin is present in the nuclei and chromosomes of A.cepa.     

Further Consideration of the Phylogeny of Some "Traditional" Heterotrichs (Protista, Ciliophora) of Uncertain Affinities, Based on New Sequences of the Small Subunit rRNA Gene

ABSTRACT. The systematic relationships and taxonomic positions of the traditional heterotrich genera Condylostentor, Climacostomum, Fabrea , Folliculina, Peritromus , and Condylostoma , as well as the licnophorid genus Licnophora , were re-examined using new data from sequences of the gene coding for small subunit ribosomal RNA. Trees constructed using distance-matrix, Bayesian inference, and maximum-parsimony methods all showed the following relationships: (1) the "traditional" heterotrichs consist of several paraphyletic groups, including the current classes Heterotrichea, Armophorea and part of the Spirotrichea; (2) the class Heterotrichea was confirmed as a monophyletic assemblage based on our analyses of 31 taxa, and the genus Peritromus was demonstrated to be a peripheral group; (3) the genus Licnophora occupied an isolated branch on one side of the deepest divergence in the subphylum Intramacronucleata and was closely affiliated with spirotrichs, armophoreans, and clevelandellids; (4) Condylostentor , a recently defined genus with several truly unique morphological features, is more closely related to Condylostoma than to Stentor ; (5) Folliculina, Eufolliculina , and Maristentor always clustered together with high bootstrap support; and (6) Climacostomum occupied a paraphyletic position distant from Fabrea , showing a close relationship with Condylostomatidae and Chattonidiidae despite of modest support.