微生物学研究中的软电离质谱技术

包括基质辅助激光解吸电离（ＭＡＬＤＩ）和电喷雾（ＥＳＩ）在内的软电离质谱是最近发展起来的质谱技术,由于这些电离方式对样品的破坏性小,质量测定范围大,分子量测定准确,样品纯度要求不高很适合分析成分复杂的微生物样品,ＭＡＬＩＤＩ－ＴＯＦ－ＭＳ结合高分辨率的二维ＳＤＳ－ＰＡＧＥ可以分析１０＾－１２摩尔水平的蛋白,是细菌蛋白质研究过程中必不可少的工具。最近的研究工作表明,通过ＭＡＩＤＩ－ＴＯＦ－ＭＳ或ＨＰ     

蛋白质组研究中肽质量指纹谱鉴定方法的建立及应用

建立了用肽质量指纹谱和数据库检索方法鉴定凝胶电泳分离蛋白南的方法。用标准蛋白质对胶上蛋白５质原位酶切制备肽谱的方法进行了讨论。分析了实际细胞蛋白质样品,获得双向电泳分离的人肺癌细胞蛋白质谱中三个蛋白质点的肽指纹谱。并通过数据７库检索分别鉴定为甘油醛－３－磷酸脱氢酶－２,测在蛋白羟基末端水解同工酶和丙糖磷酸异构酶。     

人表皮生长因子肽谱及一级结构的质谱法分析

采用液相色谱－质谱联用法（ＬＣ－ＭＳ）,分别对基因工程表达的人表皮生长因子胰蛋白酶和Ｖ８蛋白酶的酶解产物进行了肽质谱分析,获得各肽段的分子量信息。并对大部分肽段用串联质谱（ＭＳ－ＭＳ）分析了肽的序列,获得其氨基酸序列信息。此外,还测定了分子中二硫键的数目。均获得满意的结果。这一方法的建立将有助于天然的,人工合成的及基因工程表达的蛋白质或多肽的鉴定和结构分析,其精确度及分辨率都超过常规方法。     

用~1H NMR和FT－IR研究hF－GRP及其类似物TF14与DMPC脂质体的相互作用

用＾１ＨＮＭＲ和ＦＴ－ＩＲ技术研究了肽激素ｈＦ－ＧＲＰ及其类似物ＴＦ１４与ＤＭＰＣ脂质体的相互作用及其构象变化,＾１Ｈ－ＮＭＲ实验结果提示这两个肽与ＤＭＰＣ之间存在静电相互作用,这使其可从水相结合到脂相中。     

重组人骨形成蛋白-3羧基端肽在大肠杆菌中表达及其诱骨活性

以双顺反子表达载体,在大肠杆菌中经ＩＰＴＧ诱导表达了人骨形成蛋白－３羧基端肽段（ｈＢＭＰ－３Ｃ）,表达量占菌体总蛋白量的１８．５％．目的蛋白为２５ｋＤ、含ｈＢＭＰ－３Ｃ端２１５个氨基酸残基组成的肽段,包括ｈＢＭＰ－３成熟肽和一部分前肽．表达产物以包涵体的形式存在,用含ＴｒｉｔｏｎＸ－１００的洗涤液和５ｍｏｌ／Ｌ以下脲溶液连续洗涤,可获得较高纯度的重组人骨形成蛋白－３Ｃ端肽．经复性处理成可溶性蛋白,植入小鼠肌肉内,第１４ｄ组织切片显示有软骨细胞和软骨基质形成,第２１ｄ可见成骨细胞和骨基质形成．将ｒｈＢＭＰ－３Ｃ与脱矿去免疫原性异种骨粒复合后作小鼠肌肉植入试验,２１ｄ组织切片上可见硬质骨形成．结果表明：大肠杆菌表达的ｈＢＭＰ－３Ｃ经复性后具有诱骨活性,糖基化并非ＢＭＰ－３活性所必需．     

脑特异性金属硫蛋白—Ⅲ研究进展

金属硫蛋白－Ⅲ（ＭＴ－Ⅲ）是富含半胱氨酸的低分子量蛋白,主要在脑内含Ｚｎ＾２＋神经元表达,与脑的发育过程有关。ＭＴ－Ⅲ可抑制体外培养的神经细胞生长和存活。ＭＴ－Ⅲ与Ｚｎ＾２＋的结合有相对特异性,可能影响依赖于Ｚｎ＾２＋的一系列生物活动。ＡＤ患者脑内ＭＴ－Ⅲ表达减少,敲除ＭＴ－Ⅲ基因的小鼠对红藻氨酸诱发癫痫更敏感,提示ＭＴ－Ⅲ可能在某些脑变性疾病中发挥作用。     

增强UV—B辐射对蚕豆叶片微粒体膜一些性质的影响

温室条件下,用０,８．６５ｋＪｍ＾－２ｄ＾－１及１１．２ｋＪｍ＾－２ｄ＾－１不同剂量的ＵＶ－Ｂ辐射处理蚕豆幼苗。Ｃａ＾２＋－ＡＴＰａｓｅ及Ｍｇ＾２＋－ＡＴＰａｓｅ的活性在辐射处理期间下降。在处理２１ｄ,Ｔ１和Ｔ２微粒体膜的ＭＤＡ含量明显高于对照,同时ＩＵＦＡ急剧下降,且呈明显的剂量效应。     

液相色谱—电喷雾质谱法分析牛胰核糖核酸酶B酶解肽谱 …

糖蛋白分析一直是蛋白质分析鉴定的难点,为建立准确灵敏的糖蛋白分析方法。采用液相色谱－电喷雾质谱法（ＬＣ－ＥＳＩ－ＭＳ）对糖蛋白－牛胰核糖核酸酶Ｂ（ＲＮａｓｅ Ｂ）的酶解肽谱进行分析,证实其一级结构。通过比较糖苷酶处理酶解肽段前后的肽谱,确定糖基化位点,通过过串联质谱（ＭＳ／ＭＳ）解析了Ａｓｎ连接的糖型结构及去糖后肽段的氨基酸序列。糖型结构经α－甘露糖革酶处理和质谱分析确定为高甘露糖型。此外,还对糖     

木糖异构酶突变体的研究—纯化与性质

纯化了ＡｒｔｈｒｏｂａｃｔｅｒＤ－木糖异构酶的三个突变体,Ｔ８９Ｓ,Ｖ１３４Ｉ的比尖（Ｕ／ｍｇ）分别为３．４２和６．１８,Ｗ１３６Ｅ没有生。Ｔ８９Ｓ和Ｖ１３４Ｉ的活性需要二价阳离子,对于Ｔ８９Ｓ,Ｍｇ＾２＾＋的作用强于Ｃｏ＾２＾＋。在以木糖为底物时,Ｔ８９Ｓ的Ｋｍ值为４９．８ｍｍｏｌ／Ｌ,Ｖ１３４Ｉ的Ｋｍ值为９．８ｍｍｏｌ／Ｌ,在以果糖为底物时,木糖醇对Ｔ８９Ｓ是竞争性抑制剂,山梨糖醇对Ｔ８９Ｓ和     

天花粉蛋白T18肽片段的溶液构象及T14,TDK和T18肽…

用＾１Ｈ ＮＭＲ法测定Ｔ１８肽在ＤＭＳＯ（ＭＳ１８）和５０％六氟异丙醇（ＦＰ１８）中的溶液构象。ＭＳ１８含有Ｉｌｅ３￣Ｇｌｕ７和Ａｌａ１２￣Ｇｌｎ１６两段β－折叠链;而ＦＰ１８则转变为α－螺旋结构。综合分析Ｔ１４,Ｔ１８和ＴＤＫ三个模型肽的结构性质和稳定性,比较肽链序列和溶剂作用,提出肽链局部优势结构的概念,并据此讨论天花粉蛋白小结构域折叠起始过程。     

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.