银杏花粉微管蛋白的纯化及鉴定

采用丙酮粉抽提,DEAE-SephadexA-50、SephacrylS-300、MonoQ柱层析,从银杏（GinkgobilobaL．）花粉中分高纯化出微管蛋白（tubulin）,其两个亚基（α、β）的分子量分别为54kD和52kD．纯化的微管蛋白可与鸡脑微管蛋白抗体发生免疫交叉反应．     

绿豆芽中微管蛋白异型的研究

采用 DEAE-Sephadex-A50离子交换层析和 PGGE 电泳对绿豆(Phaseolus radiatus L.)芽中的微管蛋白进行分离。Western blot 和免疫点印迹分析发现了两种聚合度不同的微管蛋白异型。单体微管蛋白亚基的分子量为56kD 和56.5kD;四聚体微管蛋白亚基的分子量为54kD 和54.5kD。实验结果表明微管蛋白α、β亚基组装过程是一个相互选择的过程,这种现象可能与微管蛋白基因表达有关。     

萱草花粉类动力蛋白生物物理学及药理学性质

利用FPLC技术从萱草花粉中鉴定并纯化了动力蛋白,研究了它的酶学性质及部分生物化学性质。结果如下:纯化的类动力蛋白分子量为100kD,等电点pI=6·15和6·80。在280nm波长激发下,最大的荧光发射波长是346nm。荧光光谱分析结合紫外吸收光谱及导数光谱分析推断它含有色氨酸和酪氨酸残基。药理学性质研究表明巯基可能在酶的活性中心起重要作用。     

千里光β-微管蛋白基因结构与功能的生物信息学分析

β-微管蛋白是影响细胞新陈代谢和行使功能的重要结构物质,研究β-微管蛋白基因的序列信息对揭示其蛋白结构与功能具有重要指导意义。从千里光全长cDNA文库中分离得到β-微管蛋白基因,并采用生物信息学软件进行序列分析。结果显示,该基因长度为1750 bp,编码的蛋白质长度为448个氨基酸,与柚子β-微管蛋白的同源性最高,达96%;其蛋白质分子量为50.01 kD,理论等电点为4.83。β-微管蛋白二级结构主要组成为无规则卷曲结构和α螺旋结构;结构域分析发现该蛋白具有两个保守结构域;三级结构预测为相对稳固的类球形结构;信号肽分析将该蛋白主要定位于细胞质、过氧化物酶体、线粒体基质等亚细胞器位置。将该基因序列上传至GenBank所获得的登录号为KF887495。本实验结果为千里光β-微管蛋白的作用机制揭示和应用研究提供了基础数据,也为植物β-微管蛋白基因的分子研究提供了理论依据及基础资料。     

二化螟β1微管蛋白基因cDNA序列的克隆与序列分析

微管是真核生物体内分布最为广泛的一类蛋白,是由a-和β-两种不同的微管蛋白组成的异源二聚体.微管参与许多细胞功能,如细胞形态发生、细胞生长和分裂等.以二化螟3龄幼虫为材料提取总RNA,利用RT-PCR和cDNA末端快速扩增技术(RACE),扩增得到该虫的β微管蛋白基因的cDNA序列一条.cDNA序列含1 862个碱基,开放读码框1 344个碱基,编码氨基酸447个,分子量约为50.2kD,等电点4.82.氨基酸序列中1～4个氨基酸MREI为β微管蛋白转录后调控信号,是微管蛋白特异片段;氨基酸序列的140～146GGGTGSG位存在一个微管蛋白标志信号片段.序列比对表明,克隆的β微管蛋白基因与其他昆虫的β微管蛋白基因在核苷酸和氨基酸水平上都是高度同源的,与家蚕Bombyx moriβ1微管蛋白的氨基酸序列同源性达到99.1%,与烟草天蛾β1微管蛋白的氨基酸序列同源性达到98.7%,与果蝇Drosophila melanogasterβ1微管蛋白的氨基酸序列同源性达到96.9%.该基因cDNA序列已经登录Genbank并获得登录号为EU429675.     

调节因子IGFⅡ、TNFα、VRF促细胞增殖与β微管蛋白表达的相关性

细胞增殖必伴有染色体的一分为二及细胞质的增生,β微管蛋白则参与细胞的增殖过程．正性和负性调节因子对β微管蛋白的表达及细胞增殖间的相关性研究显示,不同生理剂量的正性调节因子ＩＧＦⅡ、Ｔ３／Ｔ４处理ＵＭＲ１０６细胞１２ｈ,Ｎｏｒｔｈｅｒｎｂｌｏｔ实验发现它们在促进细胞ＤＮＡ合成的同时,可使β微管蛋白ｍＲＮＡ表达增加,呈剂量依赖关系．而负性调节因子ＴＮＦα则相反地在抑制细胞ＤＮＡ合成的同时,使β微管蛋白ｍＲＮＡ表达降低,也呈剂量依赖关系．Ｗｅｓｔｅｒｎｂｌｏｔ实验进一步表明,ＩＧＦⅡ可使β微管蛋白表达增加,而ＴＮＦα使β微管蛋白表达降低．由此可见,β微管蛋白的合成与细胞增殖间存在着一定的相互联系．     

水稻恶苗病菌对苯并咪唑类杀菌剂抗药性分子机理研究初探

用真菌β-微管蛋白基因的丰余寡聚核着酸引物B1和B3,扩增了一段871bp的水稻恶苗病菌Fusariummoniliforme的β微管蛋白基因片段,进行了克隆和DNA序列测定,并根据该序列设计了Fmoniliformeβ-微管蛋白基因的特异性测序引物。经过对恶苗病菌对多菌灵具有不同抗性水平菌株的β-微管蛋白基因核着酸序的比较研究,表明Fmoniliforme的β微管蛋白的165,198,200和257位置氨基酸末发生突变,在克隆的片段内也未发现能引起氨基酸改变的核着酸突变。说明该菌对多菌灵产生抗性的分子机理与目前已知的其他真菌有所不同,有待进～步研究。     

萱草花粉微管蛋白的体外聚合及电镜观察

微管(microtubule)作为细胞骨架的主要成分,在植物体内,微管除决定细胞的形状外,还参与很多重要的细胞功能。但有关微管蛋白生物化学的研究绝大多数来自动物脑组织材料,对植物微管蛋白的研究除培养细胞外所知甚少,我们纯化了毫克数量的萱草(Hemer-ocallis fulvaL.)花粉微管蛋白,利用紫杉醇作为促进剂,在Mg2 、GTP等存在下体外聚合成功,并观察了其电镜下的形态。     

对虾神经系统中微管蛋白的生化和免疫荧光鉴定

采用反复解聚-聚合的方法从对虾腹神经索中分离纯化了微管蛋白。用阴离子去垢剂 SDS处理,将对虾神经纤维中的微管蛋白解聚为分子量相似的二条链。它们的分子量都在55,000左右。这与许多其他作者从各种不同组织中提取的微管蛋白的相同。以纯化的小鸡脑微管蛋白作抗原,制备了兔抗小鸡脑微管蛋白的抗体作第一抗体,以 FITC 羊抗兔抗体作第二抗体,研究了微管蛋白在对虾腹神经索中的神经纤维内的分布。令人意外的是除了对虾神经纤维特有的微管鞘与轴浆外,对虾神经纤维的髓鞘部位也呈现明显的荧光反应。这一结果提示,对虾神经纤维髓鞘中也应含有相当量的微管蛋白,但用同样方法在其它种动物(如大鼠)的有髓纤维的髓鞘中却未见阳性反应。     

甲基单胞菌Methylomonas sp.GYJ3中甲烷单加氧酶羟基化酶组分的纯化和性质

Methylomonassp．GYJ3菌株中经DEAE-SepharoseCL-6B阴离子交换层析和SephacrylS300凝胶层析分离纯化出甲烷加氧酶羟基化酶组分．经HPLC分析,纯度大于90％,分子量为240kD,纯化倍数为3．9,比活为225nmol环氧丙烷每分钟毫克蛋白．SDS-PAGE表明,羟基化酶由三个亚基组成,亚基分子量为56、43、27kD．ICPAES测定羟基化酶的Fe含量为2.1molFe每摩尔蛋白．HPLC法用于甲烷单加氧酶羟基化酶组分的纯化,纯化的羟基化酶组分比活为541nmol（环氧丙烷）每分钟毫克蛋白,是两步LC法纯化的羟基化酶的两倍,Fe含量为3．78molFe每摩尔蛋白．催化性质研究表明羟基化酶能够被化学还原剂还原为还原态羟基化酶,还原态的羟基化酶单独存在时表现出MMO活性,说明它是MMO活性中心,天然态的羟基化酶单独存在时无MMO活性,加入粗酶液中MMO活性明显增加,说明GYJ3菌中MMO是一个复合酶系．     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.