Actinobacteria的分离与鉴别

Actinobacteriaclassisnov .一般包括具有超过 50%G+C的DNA碱基组成的微生物。实验中在分土培养基中 ,添加了Nalidixicacid及Aztreonam和Benlate ,从 4份土壤样品中 ,共分离得到 64株细菌 ,革兰氏阳性细菌共计 5 6株 ,占所分离菌株的 87 5 %。任意挑选其中的革兰氏阳性细菌 ,选用以高G +C含量革兰氏阳性细菌为靶点的PHGC探针及PNHGC对照探针 ,并联合使用我们自行设计的适用于Actonobacteria     

涅斯捷连科氏菌属菌株的分离及系统学研究

从我国新疆及埃及东部高盐环境中分离到 4株形态上类似于涅斯捷连科氏菌属的革兰氏阳性菌株。以该属的 2个典型菌株 (NesterenkoniahalobiaDSM 2 0 5 4 1T和NesterenkonialacusekhoensisDSM 12 5 4 4 T)为对照 ,对 4个分离菌株进行了包括形态学 ,生长pH范围 ,温度范围 ,耐NaCl,KCl,MgCl2 ·6H2 O ,CaCl2 实验及酶学特性以及细胞化学组份、(G C)mol%含量测定 ,16SrDNA分析及DNA DNA杂交在内的多相分类研究。结果显示 ,4个分离菌株属于涅斯捷连科氏菌属 ,但与涅斯捷连科氏菌属的 2个有效发表种有显著差异 ,因此 4个分离菌株均为该属的新种     

嗜热脂肪芽孢杆菌的表观群与亲缘群的关系

用DNA-DNA体外杂交的方法考察了嗜热脂肪芽孢杆菌的表观群菌株的亲缘关系。按DNA同源性％形成三个亲缘群。亲缘群并不与表观群十分吻合,却与DNA的G+C含量更相关。同一亲缘群菌株DNA的G+c含量相近,然而DNA的。+。含量相近的并不一定属于同一亲缘群。 表观群中各菌株的中心株距离也与DNA的G十。含量相关。。州“的G+C含量接近中心株的菌株,则它与中心株距离也相对地接近。结果表明嗜热脂肪芽孢杆菌是一个异源性的种。本文结果支持Brenner的主张：将表观特征和遗传特征结台作为种的定义。     

反相高效液相色谱法测定色盐杆菌新种ST307的DNA G+C mol%含量

目的:使用反相高效液相色谱法测定色盐杆菌新种ST307的DNA G+C mol%含量.方法:以Escherichia coli DH5α为标准菌株,采用90%重蒸水10%甲醇为流动相,检测波长260nm,流速1ml·min-1,在Venusil MP C18柱上对四种碱基进行分离.结果:DNA碱基分离效果好,以外标法计算得到标准菌株DH5α的DNA G+C mol%含量为50.3%,待测菌株ST307的DNA G+C mol%含量为60.5%.结论:采用反向高效液相色谱法测定色盐杆菌的DNA G+C mol%含量准确可靠.     

双歧杆菌的数值分类及代表性菌株同源性的分析

对不同来源的55株双歧杆菌进行了数值分类研究。这些菌包括13株参照菌株和42株新分离物。在这些菌株中19株来源于人体,25株来自动物源,11株分离自污水。有的菌株是从未曾报道过的动物来源分离到的。通过数值分析比较了75项性状（形态、生理生化和对抗生素敏感性等）,以不加权平均链锁聚类的方式进行族群归类。在70％Sm水平上划分成5个聚类群和12个亚群。对这些聚类群菌株间的关系进行了分析。在数值分类树状图谱的簇群归类中,人和动物来源的菌株基本上彼此分开,而分离自污水的菌株穿插在人或动物来源的聚类群中。在数值分类的基础上对各聚类群中的一些菌株的DNA中G＋Cmol％进行了测定。依据16SrRNA可变区序列的分析,以PCR方法合成的生物素标记探针,对某些代表性菌株的DNA片段同源性进行了分析．同种的菌株间与不同种菌株间的同源性显示有差别。     

球形芽孢杆菌二元毒素基因的定位及无芽孢突变株的生物学特性

DES诱变得到了三株球形芽孢杆菌无芽孢突变株G5、C4、L5,经显微观察、超微结构分析、生物测定、蛋白质SDSPAGE分析及质粒检测,观察到突变株C4、L5阻碍于芽孢形成的第Ⅱ期,突变株G5阻碍于芽孢形成的第Ⅲ期,其细胞中已有晶体形成,它对致倦库蚊幼虫的毒力明显高于仅有二元毒素蛋白合成、而无毒素晶体形成的突变株C4和L5。突变株L5消除了质粒,仍有二元毒素蛋白合成。Southern blot分析表明含二元毒素基因部分DNA片段的探针仅与菌株C341、BS10的染色体具有同源性,因此证明供试菌株的二元毒素基因定位于染色体上。     

大连海域沉积物中可培养海洋链霉菌活性及其多样性

以大肠杆菌、金黄色葡萄球菌和尖孢镰刀枯萎病菌作为测试靶目标,采用9种分离培养基从大连海域13个不同采样点的海洋沉积物样品中分离到165株海洋链霉菌。从165株海洋放线菌中筛选到对金黄色葡萄球菌具有抑制活性的菌株85株,占总菌株数的51.5%;对大肠杆菌具有抑制活性的菌株27株,占总菌株数的16.4%;对尖孢镰刀枯萎病菌具有抑制活性的菌株仅有6株,占总菌株数的3.6%。因此,海洋链霉菌的活性更多地表现为对细菌的抗性,尤其对革兰氏阳性细菌具有更高的抑制活性。对其中具有抑制活性或形态独特的菌株进行了16S r DNA序列分析,并构建系统发育树,显示活性海洋链霉菌具有丰富的种类多样性和广谱抗菌活性。同种海洋链霉菌与土壤链霉菌活性比较结果也表明,海洋链霉菌多表现抗革兰氏阳性细菌活性。     

反相高效液相色谱法测定Bacillus claussi的DNA G+Cmol%

目的:为了建立细菌DNA G Cmol%的快速准确的测定方法,选用反相高效液相色谱法测定Bacillus claussiDNA的G C mol%。方法:以已完成全基因测序的Bacillus haloduransC-125作为标准菌株,作者实验室新分离鉴定的2株Bacillus claussi作为待测菌株。采用80 mmol/L磷酸二氢钾溶液(pH5.6)20%甲醇为移动相,检测波长260nm,流速1ml.min-1,在Kromasil C18柱上对4种碱基进行分离。结果:DNA碱基分离效果好,无杂峰干扰,以峰面积计算得到标准菌株Bacillus haloduransC-125的G C mol%为44.23%,待测菌株的G C mol%分别为45.75%、43.35%,经过统计学分析,与已经报道的G C mol%差异不显著。结论:实验结果充分显示高效反相液相色谱法测定细菌DNAG C mol%快速、准确、结果稳定,是细菌精确分类鉴定的一种可靠方法。     

黄单胞菌属的分类研究进展

引起植物病害的细菌约有300多种,广义的细菌是原核生物界中引起各种植物病害的一类最大的群体。黄单胞菌属的所有种都是植物病原细菌,引起植物病害症状多为叶斑、叶枯,少数为萎蔫、溃疡。黄单胞菌属（Xanthomonas）属于薄壁菌门、假单胞菌科,菌体短杯状,0．4－1．0×1．2－3．0μm,根极生鞭毛,严格好气性,革兰氏染色阴性,DNA中的G＋C含量为63－71mol％。细菌的分类地位和分类系统曾有过多次变动,迄今仍不完善。过去,黄单胞菌属的分类单一利用奇主的专化性划分种,即从一株寄主植物上分离到新的黄单胞菌菌株,使定为一个种,由…     

不同来源鼠李糖乳杆菌的随机扩增多态DNA分析

[目的]建立鼠李糖乳杆菌(Lactobacillus rhamnosus,Lr)菌株之间的分子鉴别方法并分析不同分离株之间的遗传多样性.[方法]从56份采集自中国新疆和田和广西巴马瑶族自治县的长寿老人粪便样本中分离得到的乳酸菌中,经生理生化分析和API 50CHL试验条鉴定,获得10株Lr.对10株Lr分离株和1株Lr标准株ATCC7469进行了随机扩增多态DNA分析,从50条随机引物中筛选到5条在菌株水平上具有鉴别力的引物P14、OPG28、OPG25、P7和P4并建立和优化了Lr菌株RAPD指纹图谱扩增方法.根据RAPD结果计算菌株间的遗传相似系数并进行聚类分析.[结果]获得了清晰稳定的DNA指纹图谱,扩增产物大小在100～2000bp之间,菌株间呈现显著的DNA多态性,不同来源的Lr分离株的遗传相似系数在0.581～0.935之间,在相似系数0.80水平上可以将11株Lr菌株分为5个类群,其中分离自新疆和田的Lr菌株归在类群B和类群C,而分离自广西巴马瑶族自治县的Lr菌株归在类群D和类群E.[结论]应用RAPD方法对Lr菌株进行分子鉴别是可行的,不同来源的Lr之间存在着较大的种内遗传多态性和不同的亲缘关系.     

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.