天然胶乳脂质的磷脂组成与分布

本文采用单向硅胶薄层色谱对“红星1号”（抗风品系）和“9－110”（抗寒品系）两种无性系天然橡胶胶乳脂质的磷脂组成进行定性分析,分离出8种磷脂组分,已检出6种,发现两种是从来没检出过的磷脂组分,此外用薄层色谱扫描和“吸光度比例系数校正法”对两种品系天然胶乳总脂,橡胶相和底层脂质中各种磷脂组分的分布进行了快速定量分析。     

高效薄层色谱测定湛江等鞭金藻培养过程脂质变化

【目的】研究湛江等鞭金藻(Isochrysis zhangjiangensis)由氮元素丰富至限制培养过程中脂质成分的变化。【方法】利用高效薄层色谱(HPTLC)分离分析微藻中脂质。【结果】随着培养基中营养物质的消耗,细胞逐渐处于胁迫状态,在这种状态下,细胞大量积累贮存脂质—甘油三脂(TAG),组成生物膜系统的单半乳糖甘油二脂(MGDG)、硫代异鼠李糖甘油二脂(SQDG)、磷脂酰甘油(PG)和磷脂酰胆碱(PC)含量降低,而游离脂肪酸(FFA)、甘油二脂(DAG)、双半乳糖甘油二脂(DGDG)、磷脂酰肌醇(PI)和磷脂酰乙醇胺(PE)则相对稳定。【结论】HPTLC可作为一种简便、可靠的微藻中脂质分离分析方法,为研究微藻油脂代谢途径以及甘油三脂(TAG)的调控积累提供有效手段。     

柱色谱硅胶的应用

色谱硅胶通常分为薄层色谱、柱色谱和液相色谱（含高效液相色谱）固定相用硅胶,它们在对物质分离、分析的机理方面基本一致。近几年由于生物技术和生物制药技术的发展,柱色谱硅胶的应用普及迅速,本文针对柱色谱硅胶的应用进行讨论与分析。     

大豆磷脂组成的研究——Ⅰ.磷脂组成的薄层色谱分析

本文采用一组酸性氯仿甲醇溶剂体系和碱性较强的硅胶薄板,经单向薄层色谱能将制备脂质体用的大豆磷脂多种磷脂组分很好地分开。在同一色谱条件下用标准磷脂进行检测,用薄层扫描进行定量,从而解决了只用微量样品(约30微克)即可直接对多种磷脂组分进行定性和定量分析的问题。     

大豆磷脂组成的研究 Ⅰ.磷脂组成的薄层色谱分析

本文采用一组酸性氯仿甲醇溶剂体系和碱性较强的硅胶薄板,经单向薄层色谱能将制备脂质体用的大豆磷脂多种磷脂组分很好地分开。在同一色谱条件下用标准磷脂进行检测,用薄层扫描进行定量,从而解决了只用微量样品(约30微克)即可直接对多种磷脂组分进行定性和定量分析的问题。     

均匀设计法优化绿色木霉菌黄色素薄层色谱展开剂系统

根据系统选择法确定了绿色木霉菌（Trichoderma viride）所产黄色素的薄层色谱展开剂组分,利用均匀设计实验及逐步回归法建立回归模型优化展开剂配比,最终得出优化的展开剂系统为为石油醚-乙酸乙酯-丙酮-乙酸=12．23：1：1．58：0．55,分离效果较理想,所得到的薄层色谱分离函数指标COF2达到了42．09±0．79．     

珠芽景天的显微与薄层色谱鉴定

运用形态解剖学和薄层色谱鉴定的方法,对植物药珠芽景天（Sedum bulbiferumMakino）进行了鉴定研究。结果显示该植物药鉴别特征明显,如茎横切面的皮层外侧及中柱薄壁组织散布有紫红色细胞,髓部细胞较小,壁不增厚;叶下表皮气孔密布,常可见2个（有时3个）相距较近;薄层色谱中具多个特征性荧光斑点。文中描述了该植物药材的性状、显微与薄层色谱特征,还与同属近缘植物药垂盆草、佛甲草、凹叶景天进行了薄层色谱比较,并结合四者茎的显微结构,探讨了相互之间的亲缘关系。研究结果为珠芽景天的应用提供了可靠的鉴定依据。     

用薄层色谱分离提纯单半乳糖和双半乳糖双甘油酯

本文选择两种溶剂体系,用两次单向薄层层析,从小麦抗寒与不抗寒品系天然橡胶胶乳分离提纯出6种单半乳糖和双半乳糖双甘油酯。并比较了它们的疏水侧链的脂肪酸组成。小麦糖脂疏水侧链脂肪酸的不饱和指数远大于天然胶乳糖脂。抗寒品系胶乳糖脂疏水侧链脂肪酸不饱和指数大于不抗寒品系。双半乳糖双甘油酯疏水侧链脂肪酸不饱和指数均大于其单半乳糖糖脂。     

大枣中氨基酸类成分的薄层色谱分析

中药大枣中含有多种氨基酸类成分,建立大枣中主要氨基酸类成分的薄层色谱鉴定方法,为大枣的相关研究提供依据。采用薄层色谱法对大枣中氨基酸类成分进行定性鉴别,本法操作简单,重复性好,特征明显,可以作为大枣中四种氨基酸的薄层色谱鉴别依据。     

冬虫夏草及杜仲磷脂成分的研究

本文对不同产地的冬虫夏草及杜仲磷脂成分进行了分析研究。以钼蓝比色法测定了它们的总磷脂含量,采用薄层色谱扫描和吸光度比例系数校正法测定了其磷脂组成及相对百分含量。冬虫夏草约含8种磷脂组分,主要成分为磷脂酰胆碱、磷脂酰叽醇、磷脂酰丝氨酸、磷脂酰乙醇胺和磷脂酸。杜仲约含6种磷脂组分,其中以溶血磷脂酰胆碱和磷脂酰胆碱为主。     

The Role of Lipids and Proteins in the Mechanism of Latex Vessel Plugging in Hevea brasiliensis

The phospholipid content of the bottom fraction of latex as well as the neutral lipids of rubber particles were determined in thirty-one clonal mother trees of RRII clones of Hevea brasiliensis Muell. Arg. Ratios between cationic proteins and anionic proteins in the B-sera, which is the sera contained in the lutoid particles in latex, obtained from clones RRIM-501, PB 6/9, RRII-105 and Tjir-l were determined by electrophoresis. The influence of these factors on plugging index (a measure of the magnitude of latex vessel plugging) was investigated. Lutoid instability, as indicated by bursting index, is negatively ocrrelated with the phospholipid content of the bottom fraction of latex. The neutral lipid content of rubber particles is positively correlated with the colloidal stability of latex. The latex vessel plugging during latex flow is found to be negatively correlated with both the lutoid stability and the neutral lipids in the rubber particle. A high ratio of cationic and anionic proteins in B-serum may also enhance the process of plugging.     

巴西橡胶树(Hevea brasiliensis)微管相关蛋白全长cDNA克隆及表达分析

从巴西橡胶树Hevea brasiliensis差减cDNA文库中分离到微管相关蛋白（Microtubule-associated protein,MAPs）基因片段,根据该基因片段序列信息,设计特异引物,采用cDNA末端快速扩增技术RACE（Rapid Amplification ofcDNA Ends）进行差异片段的5＇和3＇端的扩增,获得了长度为788bp的全长cDNA,该基因在GenBank中的登录号为AY461412.序列分析表明该基因包含完整的开放阅读框,编码144个氨基酸,与微管相关蛋白基因家族具有很高的同源性,推测该基因是微管相关蛋白基因.半定量RT-PCR检测证实它在胶乳中的表达强于叶中,胁迫处理（伤害及乙烯处理）使其表达上调.     

The lipid composition of Mycoplasma laidlawii strain B

1. Total lipid was extracted from Mycoplasma laidlawii strain B with chloroform-methanol mixtures and fractionated into neutral lipid, glycolipid and phospholipid components by chromatography on silicic acid. 2. Saponification of the glycolipid fraction, which represented nearly half of the total lipid, yielded two glycosides for which the structures O-alpha-d-glucopyranosyl-(1-->1)-d-glycerol and O-alpha-d-glucopyranosyl-(1-->2)-O-alpha-d-glucopyranosyl-(1-->1)-d-glycerol were established. 3. The ratio of monoglucosyl diglyceride to diglucosyl diglyceride increased with the age of the culture, though the total glycolipid concentration remained virtually constant. The glycolipid concentration was unaffected by the addition of cholesterol to the culture medium. 4. The phospholipid fraction consisted of two components, phosphatidylglucose and phosphatidylglycerol. Organisms harvested at acidic pH also contained O-amino acyl esters of phosphatidylglycerol. No lipids containing inositol could be detected.     

Comparative Study on Plant Latex Particles and Latex Coagulation in Ficus benjamina,Campanula glomerata and Three Euphorbia species

Among latex-producing plants, mainly the latex of Hevea brasiliensis has been studied in detail so far, while comprehensive comparative studies of latex coagulation mechanisms among the more than 20,000 latex-bearing plant species are lacking. In order to give new insights into the potential variety of coagulation mechanisms, the untreated natural latices of five latex-bearing plants from the families Euphorbiaceae, Moraceae and Campanulaceae were visualised using Cryo-SEM and their particle size compared using the laser diffraction method. Additionally, the laticifers of these plants species were examined in planta via Cryo-SEM. Similar latex particle sizes and shape were found in Ficus benjamina and Hevea brasiliensis. Hence, and due to other similarities, we hypothesize comparable, mainly chemical, coagulation mechanisms in these two species, whereas a physical coagulation mechanism is proposed for the latex of Euphorbia spp. The latter mechanism is based on the huge amount of densely packed particles that after evaporation of water build a large surface area, which accelerates the coagulation procedure.     

Identification and comparison of natural rubber from two Lactuca species

Renewed interest in the identification of alternative sources of natural rubber to Hevea brasiliensis has focused on the Compositae family. In our search for Compositae models for rubber synthesis, we extracted latex from stems of two lettuce species: Lactuca serriola, prickly lettuce, and Lactuca sativa cv. Salinas, crisphead lettuce. Both species contained cis-1,4-polyisoprene rubber in the dichloromethane-soluble portions of their latex, and sesquiterpene lactones in their acetone-soluble portions. The rubber from both species and their progeny had molecular weights in excess of 1,000,000g/mol, and polydispersity values of 1.1. Rubber transferase activity was detected across a range of farnesyl diphosphate initiator concentrations, with decreased activity as initiator concentrations exceeded putative saturation. These results add lettuce to the short list of plant species that produce high molecular weight rubber in their latex. Due to the genomic and agronomic resources available in lettuce species, they provide the opportunity for further dissection of natural rubber biosynthesis in plants.     

A role for a Hevea latex lectin-like protein in mediating rubber particle aggregation and latex coagulation

An in vitro aggregation of washed lutoid membrane and rubber particles, respectively, prepared from the bottom (lutoid) fraction and rubber layer of centrifuged fresh latex, leading to the formation of rubber coagulum necessary for a latex coagulation was demonstrated. A Triton X-100 extract of washed lutoid membrane proteins, isolated and prepared from the bottom fraction of centrifuged fresh latex was examined for its role in the latex coagulation process. It induced agglutination of rabbit erythrocytes, indicating the presence of a lectin-like protein. Hevea latex lectin-like protein (HLL) was purified to homogeneity by active chitin binding separation, followed by DEAE-Sepharose chromatography. Its M(r) analyzed by SDS-PAGE was 17 kDa, whereas that determined by gel filtration was 267 kDa. The HLL had a pI value of 7.2. Several glycoproteins were shown to inhibit the HLL-induced hemagglutination. The hemagglutinin activity of HLL was enhanced by Ca(2+). Of most interest was the finding that HLL strongly induced aggregation of the Hevea latex rubber particles (RP). This strong RP aggregation leads to latex coagulation, indicating the possibility that it is involved in the formation of the coagulum that plugs the latex vessel ends and stops the flow of latex upon tapping. In addition, the purified HLL also induced aggregation of RP taken from several other non-Hevea latex producing plants. This might indicate either a common or universal role of this lectin-like protein in RP aggregation and hence latex coagulation. This paper, for the first time, provides clear and unequivocal evidence for either a key biological role or physiological function of an endogenous latex lectin-like protein in the sequential process of latex coagulation.     

橡胶树死皮病胶乳C-乳清差异表达蛋白质的筛选与鉴定

橡胶树死皮病(Tapping Panel Dryness,TPD)在世界各橡胶种植园普遍发生,给橡胶种植业带来严重的危害。为了更好地了解和阐明死皮病发生、发展的分子机制,本研究应用双向凝胶电泳技术（2-DE）比较橡胶树死皮株与健康株胶乳C-乳清蛋白质组表达的差异。采用固相pH梯度双向凝胶电泳分离橡胶树死皮株与健康株C-乳清的总蛋白质,凝胶经考染显色后,用PDQuest7.40图像分析软件进行比较分析,识别差异表达的蛋白质。这些点经过胶内酶切后进行基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)分析获取肽质指纹图谱(PMF),Mascot软件搜索SWISS-PROT和NCBInr数据库鉴定蛋白质。结果：①橡胶树死皮株与健康株C-乳清凝胶的平均蛋白质点数分别为1075±35和1134±27,其平均匹配的点数分别为982±38和1008±22,组内图像匹配率达91.89﹪和88.72﹪。②橡胶树死皮株与健康株C-乳清组间的平均匹配蛋白点数为970±25。利用MALDI-TOF-MS质谱技术对40个差异明显的蛋白点进行分析鉴定,通过查询数据库鉴定了27个蛋白质。本研究建立了分辨率高且重复性较好的橡胶树死皮株与 健康株胶乳C-乳清的双向凝胶电泳图谱,并应用质谱技术鉴定了其中表达差异的蛋白质点,这些差异表达的蛋白质可能参与了死皮发生和发展的过程。     

Hev b 9, an enolase and a new cross-reactive allergen from hevea latex and molds. Purification, characterization, cloning and expression.

Natural rubber latex allergy is an IgE-mediated disease that is caused by proteins that elute from commercial latex products. A complementary DNA (cDNA) coding for Hev b 9, an enolase (2-phospho-D-glycerate hydrolyase) and allergen from latex of the rubber tree Hevea brasiliensis, was amplified by PCR. The PCR primers were designed according to conserved regions of enolases from plants. The obtained cDNA amplification product consisted of 1651 bp and encoded a protein of 445 amino-acid residues with a calculated molecular mass of 47.6 kDa. Sequence comparisons revealed high similarities of the Hevea latex enolase to mold enolases that have been identified as important allergens. In addition, the crucial amino-acid residues that participate in the formation of the catalytic site and the Mg2+ binding site of enolases were also conserved. Hevea latex enolase was produced as a recombinant protein in Escherichia coli with an N-terminal hexahistidyl tag, and purified by affinity chromatography. The yield amounted to 110 mg of purified Hev b 9 per litre of bacterial culture. The recombinant allergen bound IgE from latex, as well as mold-allergic patients, in immunoblot and ELISA experiments. The natural enolase was isolated from Hevea latex by (NH4)2SO4 precipitation and ion exchange chromatography. The natural and the recombinant (r)Hev b 9 showed equivalent enzymatic activity. Patients' IgE-antibodies preincubated with rHev b 9 lost their ability to bind to natural (n) Hev b 9, indicating the identity of the B-cell epitopes on both molecules. Cross-reactivity with two enolases from Cladosporium herbarum and Alternaria alternata was determined by inhibition of IgE-binding to these enolases by rHev b 9. Therefore, enolases may represent another class of highly conserved enzymes with allergenic potentials.     

The isolation and composition of helical protein microfibrils from Hevea brasiliensis latex

1. The microfibrils contained within the lutoid particles of Hevea brasiliensis latex obtained from young tissue have been isolated by methods based on low-speed centrifugation, isoelectric precipitation and gel filtration. 2. The isolated microfibrils behave as a single protein having an isoelectric point of about 4 as determined by paper electrophoresis. 3. The only components so far detected in the microfibrils are protein and possibly carbohydrate; nucleic acid appears to be absent. 4. The amino acid composition of the microfibril protein shows no unusual features. 5. In latex from the more mature laticiferous tissues of H. brasiliensis, the lutoid particles appear to be devoid of microfibrils or their protein decomposition products.     

Comparing the content of lipids derived from the eye lenses of various species

The lipid content in the eye lens was analyzed and compared among various species in this study. The eye lens lipids of the following species were investigated: cow, horse, duck, and freshwater trout. Additionally, the lipids derived from cataractous bovine lens and from cataractous human eye lens lipoprotein complexes were analyzed. The following lipid classes were detected in clear lenses: cholesterol, sphingomyelin, phosphatidylcholine, phosphatidyletanolamine, and phosphatidylserine. In cataractous bovine lens and in lipoprotein complexes from human nuclear cataract, phosphatidyloinositol and phosphatidyloglycerol were detected. Cholesterol and sphingomyelin, essential for hypothetical formation of cholesterol-rich domains, were the most abundant lipids in the lenses of all investigated species. These two components of eye lens lipid fraction were analyzed quantitatively using thin layer chromatography and spectrophotometric assay; the other lipids were identified qualitatively using thin layer chromatography.