Frankia基因组DNA提取及其PCR带型

本文采用溶菌酶法、CTAB法、微波（MW）法及CTAB＋MW法等4种方法提取Frankia菌CpIl基因组DNA。结果表明四种方式均可满足试验要求,其中以CTAB法及微波法最为有效。在比较提取方法与PCR带型间的关系中,发现提取上的差异不影响PCR带型的变化。     

ISOLATION AND CHARACTERIZATION OF CHROMATIN FROM NEUROSPORA CRASSA

Different preparations of chromatin isolated from mycelia of Neurospora crassa were analyzed for DNA-associated RNA and proteins. The UV absorption spectra, the ultrastructure of chromatin, and the amino acid composition of the acid-extractable proteins were studied. The protein:DNA ratios range from 1.5 to 2.8; the RNA:DNA ratios range from 0.5 to 1.24. UV absorption shows a macimum at 259 mµ and a minimum at 238–239 mµ. The E280/E260 ranges from 0.59 to 0.70. Electron microscopy reveals a fibrous structure with individual fibers of 120–150 A average diameter. Attempts were made to study the protein by polyacrylamide gel electrophoresis and amino acid analysis. The results indicate that Neurospora chromatin does not contain basic proteins comparable to calf thymus histone. The ratios of basic to acidic amino acids range from 0.93 to 1.19. On electrophoresis, no bands are seen whose positions correspond to those of histones. Staining for basic proteins with fast green or eosin Y at pH 8.2 also shows a negative reaction, suggesting the absence of histones.     

四棱豆根瘤菌的分离及特性

从四棱豆根瘤上分离出一株慢生型根瘤菌(Rhizolia Ps)。在显微镜下,该菌株为革兰氏阴性,有一根亚极生鞭毛,大小为0.59×1.49微米。其增代时间为15.52小时,在39℃和含1.5％NaCl培养基中均不能生长。回接时能使四棱豆幼苗根部结瘤,固氮酶活性为2.81微摩尔乙烯·克鲜瘤~(-1)·小时~(-1)。本文对该菌株的含碳化合物利用、B.T.B.反应、在肉膏蛋白胨上的生长情况、石蕊牛奶反应、淀粉和明胶水解等生理生化特征进行了试验。     

桃金娘中可水解丹宁的分离与结构

利用Diaion HP-20、Toyopearl HW-40及MCI gel CHP-20P等反复柱层析,从桃金娘(Rhodomyrtus tomentosa(Ait)Hassk)叶及根中分得4种可水解丹宁,其中3种为C-甙可水解丹宁。经化学方法和波谱分析,分别确定为:pedunculagin(Rt-2),casuariin(Rt-3),castalagin(Rt-9)和一新的C-甙可水解丹宁(Rt-8),命名为山稔甲素(tomentosin),这些化合物均为首次从该植物中得到。     

肉苁蓉中苯乙醇甙类成分的分离和鉴定

从列当科药用植物肉苁蓉(Cistanche deserticola)中分得四个苯乙醇甙类成分,经光谱分析,确定这四个成分是海胆甙(echinacoside),肉苁蓉甙A(cistanoside A),麦角甾甙(acteoside)和2＇-乙酰麦角甾甙(2＇-acetylacteoside)。以上成分在该植物中属首次分得。     

ISOLATION AND CHARACTERIZATION OF LUMINAL MEMBRANES FROM URINARY BLADDER

A method is reported for the isolation of a highly purified fraction of urinary bladder membranes containing hexagonal plaques. The method uses zonal centrifugation as the final step of fractionation. The purified fraction was characterized by its electron microscopic morphology, by its enzymatic profile, by quantitative and qualitative analysis of lipids and by the protein pattern obtained by electrophoresis in polyacrylamide sodium dodecyl sulfate gels. The fraction contains 65% lipids and 35% proteins. The major protein component has a molecular weight of 27,000 daltons. Phospholipids are more than the 54% of the total lipid weight. Phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol are the major phospholipids with 50%, 30%, and 7% of the total lipid phosphorus, respectively. The glycolipid fraction is 10% of the total lipid weight and is formed by only two components, both sulfatides. Total cholesterol makes up 36% of the total neutral lipid fraction of which cholesterol esters constitute 6%. Glycoproteins are also found to be present in the fraction.     

胡萝卜(Daucus carota L.)胚性细胞蛋白的分离研究

应用IEF/SDS-PAGE双向电泳技术,比较了胡萝卜胚性细胞、非胚性细胞和不同发育时期的胚状体中可溶性蛋白的双向电泳图谱,结果发现在胚性细胞中特异存在的胚性细胞蛋白在不同发育时期的胚状体中也存在,但在非胚性细胞中不存在。因此,推测体细胞胚胎发生所需的一些基因在胚性细胞中就早已表达了。我们还成功地分离和测定了ECP 45-2 N-末端和中央部分氨基酸序列。与已知氨基酸序列相比,ECP 45-2部分氨基酸序列与ECP 45-1 部分氨基酸序列具有较高比例的同源性。因此, ECP 45-1和45-2可能属于同一因基家族。     

中华绒螯蟹组织中一株类志贺邻单胞菌的分离与特性分析

从江苏省启东市某水产养殖场患病的中华绒螯蟹组织中,分离到一株类志贺邻单胞菌,定名为JE－1株。该菌株为革兰阴性短杆菌,具端生单鞭毛,主要生化特性为氧化酶阳性,发酵葡萄糖产酸不产气,发酵麦芽糖,乳糖,肌醇等。不发酵甘露醇,鼠李糖,阿拉伯糖等;触酶,精氨酸双水解酶,鸟氨酸脱羧酶,赖氨酸脱羧酶阳性,对小鼠,鹌鹑,鲫鱼血细胞无溶血性,最适生长条件为温度28℃,pH7,含1％NaCl,经健康蟹回感,未见发病。     

桑白皮中壳聚糖的分离与鉴定

首次报道了一种从桑白皮中分离壳聚糖的简便方便。经碱醇液高温处理 5h,壳聚糖收率为 7.2 % ,游离氨基为 2 9.6% ,脱乙酰度为 2 5% ,1%浓度壳聚糖的 1%醋酸溶液粘度为 1.52 MPa·s,平均分子量为 1.3× 10 3。     

健康动物肠道正常菌群的分离与鉴定

本文采用多种选择培养基,从健康动物猪、鸿、兔、鱼肠道中分离培养出２２株正常菌群,经过菌落形状、革兰染色、美兰染色镜检及生化反应,初步鉴定为链球菌８株、孔杆菌２株、双歧杆菌４株、芽胞杆菌５林、酵母菌３株。此为优选菌种、制备微囊及生产生态制剂等奠定了良好的基础．     

花粉生殖细胞的大量分离与纯化

被子植物的花粉生殖细胞是精子的前体,在生殖过程中占有重要地位。由于它被营养细胞与花粉壁包围,一般只能以花粉粒为单位进行研究。1986年以来,我们用压片法分离出花粉生殖细胞,作了有关细胞生物学研究,同时指出分离的生殖细胞用于离体培养与遗传操作的意义。作者参考了 Russell 分离精子的一步“渗透压冲击法”,建立了适于大量分离与纯化生殖细胞的“二步渗透压冲击法”(以下简称“二步法”)。此外,还作     

海芋胰蛋白酶抑制剂的分离纯化及性质研究

利用亲和层析和分子筛凝胶过滤等技术,从海芋根茎中分离纯化到一种胰蛋白酶抑制剂,简称ＡＭＴＩ。经ＰＡＧＥ、ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎ ｂｌｏｔ鉴定均显示单一条带,经ＳＤＳ－ＰＡＧＥ测定,其分子量为２２０００,经等电聚焦（ＩＥＦ）测定,其等电点为６．２。根据对胰蛋白酶的抑制比可知该抑制剂为单头抑制剂,其抑制活性在６０℃和ｐＨ５￣１１范围内保持稳定。     

ISOLATION AND FLOW CYTOMETRIC CHARACTERIZATION OF PROTOPLASTS FROM MARINE MACROALGAE

Protoplasts were isolated from Ulva rigida C. Agardh (Chlorophyta) and two species of Rhodophyta , Gracilariopsis lemaneiformis ( Bory) Dawson, Acleto et Folvik and Gracilaria tenuistipitata Chang et Xia var . liui with minor modifications (the inclusion of 0.01% agarase in the set of cell-wall-degrading enzymes for the two red algae). Flow cytometric characteristics of freshly isolated protoplasts were determined on a FACScan flow cytometer (FC). The most useful parameters for characterizing protoplasts from marine algae were forward angle light scatter (FSC), orange fluorescence (FL2) and red fluorescence (FL3). Protoplasts from all the species were easily distinguishable when their FSC, FL2, and FL3 signals were combined in the bivariate plots FL3 vs. FSC and FL3 us. FL2. Two alternative techniques to help identify protoplasts from debris in the FC computer screen were developed (for FC without sorting capability). Both techniques were based on the ability of new FCs to record time. The first one was based on the induction of rapid changes of cell volume in response to osmotic stress. Only intact protoplasts responded to changes in the osmotic pressure. The second one was based on the uptake and hydrolysis of fluorescein diacetate by intracellular esterases. Viable protoplasts showed a hyperbolic accumulation of fluorescein with time. Semimaximal fluorescein accumulation was attained in 30.5 ± 9.5 s. Debris was easily recognized since, contrary to protoplasts, it did not show a time-dependent accumulation of fluorescein .     

小定鞭藻毒素的分离与鉴定

从大量死鱼的鱼池中收集分离出小定鞭藻Ｐｒｙｍｎｅｓｉｕｍｐｏｒｖｕｍ的毒株,并在实验室成功地进行了单种培养,当温度２３℃,光照６００－８００ｌｘ盐度约１２—１６‰左右时,该藻在海水及人工海水培养基中均生长良好,在对数生长末期到平衡期溶血毒素活性最高。从藻细胞及浓缩的培养液中提取出二种毒素：溶血毒素（Ｈａｅｍｏｌｙｔｉｃｔｏｘｉｎｓ）和鱼毒素（Ｉｃｈｔｈｙｏｔｏｘｉｎｓ）。用新鲜牛血球测定了溶血毒素活性;用孔雀鱼测定了鱼毒素活性。用部分纯化的溶血毒素经元素分析、红外光谱、核磁共振及ＦＡＢ质谱测定,结果显示该藻溶血毒素可能是一个糖脂。     

长梗千里光中的呋喃雅槛兰型倍半萜

长梗千里光（ＳｅｎｅｃｉｏｋａｓｃｈｋａｒｏｖｉｉＣ．Ｗｉｎｋｌ．）为菊科千里光属的多年生草本植物,广布于甘肃省东南部的灌木丛中及林边草地,四川、青海也有分布［１］。其化学成分未见报道。千里光属植物的主要成分是双稠吡咯啶生物碱和呋喃雅槛兰型倍半萜［２,３］。前者具有抗癌活性,但也有致癌、致畸等毒性［４］。我们从长梗千里光的地上部分分离到两个呋喃雅槛兰型倍半萜（１）和（２）,通过ＵＶ、ＩＲ、ＭＳ、１Ｈ－ＮＭＲ和１３Ｃ－ＮＭＲ等光谱分析,确定化合物１为１α－乙酰氧基－６β－当归酰氧基－１０β－羟基－…     

红曲菌代谢产物中低极性组分的分离及表征

以石油醚∶醋酸乙酯 =4∶1(V/V)作为洗脱剂 ,采用柱层层析粗分离醇溶性红曲菌代谢产物中的低极性组分。经浓缩、结晶除去白色结晶后的浓缩液 ,用正己烷∶醋酸乙酯 =9∶1(V/V)作为展开剂进行薄层层析分离 ,在紫外灯下观察 ,从低极性组分中分离出六个组分 ,分别为 :具荧光组分、两个相隔较近的黄色组分、淡黄色具荧光组分、具浅蓝绿色荧光组分、具荧光组分。各组分的Rf 值分别为 :0 2 9、0 15、0 12、0 0 9、0 0 6、0 0 4。MS测定结果表明 ,Rf 值最大的具荧光组分可能为含有 OH及Br的共轭烯烃或脂肪酮 ,而在紫外灯下呈淡黄色组分为含有 OH的环状化合物。     

不同时期鸡胚原始生殖细胞分离的研究

采用Ficoll密度梯度离心,酶解离两种方法在鸡胚孵化的第14期、19期、28期,分离、培养鸡胚中的原始生殖细胞(PGCs)。探索PGCs分离、培养的适宜时期及方法,以期获得较多数量,较高活力的PGCs作介导生产转基因鸡。结果表明：1．提取、分离PGCs的最佳时期依次为19期、28期。2．两种分离方法均能分离到一定数量的PGCs细胞。但在19期和28期,酶解离法分离到的PGCs的相对数量较多,存活时间较长,是一种较适宜的分离方法。     

ISOLATION AND CHARACTERIZATION OF GLUCOCEREBROSIDES FROM ADULT RAT BRAIN1

Abstract— By chromatography on borate-coated silicic acid, glucocerebrosides, galactocerebrosides, sulfatides and sphingomyelins from brain tissue could be efficiently separated. Adult rat brain was found to contain 54.1 ± 1.5 nmol of glucocerebrosides per gram fresh weight. Ninety percent of the glucocere-broside fatty acids were palmitate, stearate and oleate; fatty acids with chain lengths above C₂₀ were virtually absent. No hydroxy fatty acids were found. The long chain bases of adult rat brain glucocerebrosides consisted of 74.6% C₁₈-sphingosine, 24.4% C₁₈-sphinganine and 1.1% C₂₀-sphingosine. These results are compared to those obtained from glucocerebrosides from immature rat brains (Abe & Norton , 1974) and discussed in respect to changes occurring during brain development.