WL营养琼脂对葡萄酒相关酵母的鉴定效果验证

利用WL营养琼脂对采自葡萄园和葡萄汁发酵过程中的35株酵母菌进行了分类鉴定,同时进行了5.8S-ITS和26S rDNA D1/D2区的扩增与测序。结果表明利用WL营养琼脂的鉴定结果与测序结果基本符合。WL营养琼脂是一种较为有效的葡萄酒相关酵母菌的分类鉴定培养基。     

酵母样真菌快速分型诊断

为进一步解决感染酵母样真菌病患者的早期诊断和治疗,我们采用了一种快速,简便,准确鉴定和分类酵母样真菌的方法－纸片法,同时对纸片法与传统的方法（生长图谱法）进行了比较。２１７株酵母样真菌的鉴定结果准确率达１００％,其中白念珠菌检出率最高１５５株占７１．４２％,为最常见菌,其次是热带念珠２０株占９．２２％,其它四种念珠菌１６株占７．３７％,还有分类及种未定酵母菌２６株占１１．９８％。以上观察结果其检出     

胀罐酱油中耐渗透压酵母菌分离及鉴定

从广东某调味食品厂的3批次的胀罐酱油中分离到3株引起胀罐的耐渗透压的酵母菌,这些酵母菌在含有50％～60％葡萄糖和15％NaCl的培养基上生长较快,在普通酵母培养基上生长缓慢。通过对其进行形态特征观察和生理生化的测定,初步鉴定这3株酵母菌为埃切毕赤酵母（Pichia etchellsii）。     

酵母样真菌快速分型诊断

为进一步解决感染酵母样真菌病患者的早期诊断和治疗,我们采用了一种快速、简便、准确鉴定和分类酵母样真菌的方法——纸片法,同时对纸片法与传统的方法(生长图谱法)进行了比较。217株酵母样真菌的鉴定结果准确率达100％,其中白念珠菌检出率最高155株占71.42％,为最常见菌,其次是热带念珠20株占9.22％,其它四种念珠菌16株占7.37％,还有分类及种未定酵母菌26株占11.98％。以上观察结果其检出率和国内外报道的资料相似。我们认为用纸片法鉴定酵母样真菌,操作过程简单易行、费用低廉、准确快速,可尽早为临床提供治疗依据。     

BD Phoenix^TM酵母菌鉴定板对临床分离酵母菌鉴定能力的评估

目的 评估BD Phoenix^TM酵母菌鉴定板对酵母菌的鉴定能力.方法 选取白念珠菌18株,热带念珠菌22株,光滑念珠菌19株,克柔念珠菌8株,近平滑念珠菌20株,新生隐球菌14株,季也蒙念珠菌4株,平常念珠菌1株,葡萄牙念珠菌1株,头状地霉2株,挪威念珠菌1株,链状念珠菌1株,乳酒念珠菌1株,希木龙念珠菌1株,解脂念珠菌3株,皱褶念珠菌1株,菌膜念珠菌3株,共计120株.借助Phoenix^TM 100全自动微生物鉴定仪,使用BD Phoenix^TM酵母菌鉴定板鉴定上述菌株.用真菌通用引物ITS1与ITS4对所有受试菌株的rDNA进行PCR扩增,对PCR产物进行序列测定、分析并作为金标准与BD Phoenix^TM酵母菌鉴定板的结果比较,同时使用MALDI-TOF MS质谱分析对试验菌株进行菌种鉴定.结果 BDPhoenix^TM酵母菌鉴定板除了对1株挪威念珠菌、1株平常念珠菌、1株解脂念珠菌、1株皱褶念珠菌未能鉴定以及1株链状念珠菌、1株克柔念珠菌、2株菌膜念珠菌、1株解脂念珠菌鉴定错误外其余试验菌株鉴定均正确,鉴定准确率为92.5％.所有鉴定结果均在17 h内获得,而白念珠菌、热带念珠菌、近平滑念珠菌的鉴定时间均小于6h,并且不受推荐培养基的限制.所有菌株MALDI-TOF MS的鉴定结果与其rDNA ITS序列分析的结果完全一致.结论 BD Phoenix^TM酵母菌鉴定板对多数酵母菌能够快速准确地鉴定到种,但对某些少见酵母菌的鉴定能力有待进一步考证.     

斑马鱼肠道中一株红酵母(Rhodotorula)的分离与鉴定

目的:分离及鉴定来自于斑马鱼肠道中的酵母菌.方法:采用马铃薯葡萄糖培养基( PDA),从斑马鱼肠道中分离到1株酵母菌,编号为ZF -2,进行形态学观察、生理特征、PCR扩增26S rDNA D1/D2区以及基因测序分析,并构建系统发育进化树.结果:ZF -2分离株为粉红色菌落、菌体呈椭圆形,芽殖;可以同化利用葡萄糖、乳糖、蔗糖等糖类;PCR扩增获得26S rDNA D1/D2区序列长度为642bp( HQ323251),序列比对分析表明与斯鲁菲亚红酵母(Rhodotorula slooffiae)相似性在99％-100％之间,构建的系统发育进化树显示菌株ZF-2与R.slooffiae( EU583485)关系最近,且位于同一分支.结论:首次从斑马鱼肠道中分离到斯鲁菲亚红酵母.     

芝麻香型白酒大曲中酵母菌的分离与鉴定

为了开发利用白酒大曲中的酵母菌资源,采用稀释涂布平板法,从芝麻香型白酒大曲中分离得到15株酵母菌株。利用26S rRNA基因序列分析技术对其进行分类鉴定。结果表明,其中6株酵母菌为酿酒酵母（Saccharomyces cerevisiae）,6株为库德里阿兹威氏毕赤酵母（Pichia kudriavzevii）,3株为热带假丝酵母（Candida tropicalis）;并对代表菌株Y1、Y2及Y4进行了形态观察;最后通过随机扩增多态性DNA标记（random amplified polymorphic DNA, RAPD）对分离得到的酵母菌进行分型,表明6株酿酒酵母分属于5个株型,6株库德里阿兹威氏毕赤酵母分属于5个株型,3株热带假丝酵母分属于3个株型。     

耐高温、耐酸产酒精酵母的筛选与鉴定

以白酒厂窖泥和生产用酒曲为材料,富集、分离得到32株酵母菌,从中筛选到1株产乙醇能力强,耐高温、耐强酸、起酵速度快的菌株Y30。该菌株能在45℃正常发酵,且致死温度达到59℃,比文献中报道高5℃;能在pH2．5的条件下正常发酵;能耐14％的酒精度;起酵速度快,接种4h后即开始发酵。对菌株进行形态分析、生理生化实验、Biolog鉴定和分子鉴定,确定该菌株为Saccharomyces cerevisia。该菌株的成功选育为后续高效生产乙醇提高了很好的菌种来源。     

临床标本中酵母样真菌的分离鉴定和药敏结果分析

目的调查目前临床标本中酵母样真菌感染现状及耐药性情况。方法常规分离临床标本中的酵母样真菌,用科玛嘉显色培养基及法国生物-梅里埃API20CAUX鉴定系统鉴定菌种,并用K—B法进行药敏试验。结果研究期间共分离到193例有临床意义的酵母样真菌,检出率较高的依次为白假丝酵母菌（62．7％）,热带假丝酵母菌（20．2％）,光滑球拟酵母菌（9．3％）。在6种抗真菌药中,对伊曲康唑、氟康唑的耐药率增高。结论近年来由酵母样真菌感染引起的疾病及其对常用抗真菌药物的耐药率有明显增加,应加强真菌的检测和耐药性监测工作,以指导临床合理使用药物。     

固囊酵母的一个新种

从中国保定槐茂甜面酱中分离到一株酵母。经鉴定,这株酵母菌属于固囊酵母属(Citeromyces),并为该属中的一个新种,命名为保定团囊酵母(Citeromyces baodingensis zhangsp.Nov.)。保定团囊酵母与固囊酵母生理生化特性有显著区别,保定固囊酵母(Citeromycesbaodingensis)不发酵蔗糖、棉子糖,同化半乳糖和纤维二糖,不同化海藻糖和棉子糖,G+C含量为48.5mol％。     

酵母菌快速鉴定新方法:用DHPA法及ID 32C试条鉴定酵母菌

目的：建立酵母菌快速鉴定新方法。方法：依据双歧鉴定法和层次分析原理建立DHPA法，用DHPA法及ID32C试条鉴定酵母菌。结果：DHPA法和32C法对酵母菌63个KIU鉴定的正确性均为100％。DHPA法与32C法对酵母菌3759个试验编码分析的总符合率为81．5％。结论：DHPA法解决了32C法存在的缺码或不能鉴定问题，是酵母菌准确、快速、简便、经济和实用的鉴定方法。     

RESPONSIVENESS OF YEAST CELLS TO AUXIN, ANIMAL SEX HORMONES AND YEAST SEXUAL HORMONES IN RELATION TO SEX CONTROLLING GENES

Genetic control of the ability of yeast cells to respond to cell-expanding action of animal, plant and yeast hormones was studied. Mating type specificity in the ability to respond to yeast sexual hormones was changed by introduction of homothallism-controlling genes, D and HM. Auxin, a plant hormone can expand cells of most homothallic strains, but not those of heterothallic strains as far as tested. Animal sex hormones showed the action similar to that of yeast sexual hormones.     

酵母菌中超氧化物歧化酶的研究

首次提出了用甲苯破壁提取酵母菌中SOD的方法,此法提取SOD的含量分别为氯仿-乙醇法、酶裂解法和细胞自溶法的1.29、1.08和2.01倍.通过对5种酵母菌的测定,发现酵母菌对氧的抗性与其SOD含量密切相关;同时对一株SOD含量较高的卡尔酵母(Saccharomyces carlsbergensis)BY2菌株进行了营养和培养条件的研究,结果表明它们对该菌株SOD含量和SOD同功酶的影响较大.     

猪源双歧杆菌的分离与鉴定

运用双歧杆菌选择性培养基,从１～４周龄乳猪的粪便中共分离纯化到５２个菌株。通过染色镜检、生化反应、代谢产物分析、抗生素敏感性试验研究表明这些菌株均与双歧杆菌属特征相符。根据糖发酵试验初步鉴定结果,其中４５个菌株为小猪双歧杆菌,５株为猪双歧杆菌,２株为嗜热双歧杆菌。部分菌株的急性毒性试验表明受试菌株对小白鼠无任何毒性副反应。     

GENETIC APPROACHES TO THE IDENTIFICATION OF INTERACTIONS BETWEEN MEMBRANE PROTEINS IN YEAST

ABSTRACT

The recent sequencing of entire eukaryotic genomes has renewed the interest in identifying and characterizing all gene products that are expressed in a given organism. The characterization of unknown gene products is facilitated by the knowledge of its binding partners. Thus, a novel protein may be classified by identifying previously characterized proteins that interact with it. If such an approach is carried out on a large scale, it may allow the rapid characterization of the thousands of predicted open reading frames identified by recent sequencing projects. Currently, the yeast two-hybrid system is the most widely used genetic assay for the detection of protein–protein interactions. The yeast two-hybrid system has become popular because it requires little individual optimization and because, as compared to conventional biochemical methods, the identification and characterization of protein–protein interactions can be completed in a relatively short time span. In this review, we briefly discuss the yeast two-hybrid system and its application to large scale screening studies that aim at deciphering all protein–protein interactions taking place in a given cell type or organism. We then focus on a class of proteins that is unsuitable for conventional yeast two-hybrid systems, namely integral membrane proteins and membrane-associated proteins, and describe several novel genetic systems that combine the advantages of the yeast two-hybrid system with the potential to identify interaction partners of membrane-associated proteins in their natural setting.     

海南岛两个虫媒病毒分离物的初步鉴定

本文报告,1985年夏从海南岛采集的6种蚊子中,分离到4株乙型脑炎病毒和38株非乙脑病毒,其中两株(HN8和HN99)为甲组虫媒病毒。二者抗原性非常相近,对乳小白鼠脑内、腹腔和皮下接种部有较强的致病力,能在多种组织培养细胞上增殖并产生病变,能在鸡胚纤维母细胞上形成空斑,能凝集多种动物红细胞,血凝pH范围为5.75～6.4,最适pH为5.75或6.0,血凝素在4～37℃均较稳定,血凝素对福尔马林敏感。病毒呈球形,有包膜,直径60～65nm,属RNA病毒,对热、酸、乙醚和紫外线均敏感。血清学鉴定表明这两株病毒只与甲组虫媒病毒抗体有反应,尤其与MAY病毒抗原性关系较密切。同年从当地人、畜血清中检查了HN99病毒抗体,其阳性率为:健康人14％,猪23％,山羊35％。另外还与北京的M4株病毒作了初步比较,并对新毒株及我国其它有关毒株的分类学地位提出了初步看法。     

酵母(Sacchar omyces cerevisiae)的PFG电泳核型比较分析

我们用脉冲式梯度电场凝胶电泳技术,对一些啤酒酵母菌进行了电泳核型分析,比较了不同菌的染色体DNA电泳核型的异同。通过用染色体专一探针做Southern杂交,找出第Ⅰ—ⅩⅥ染色体DNA在PFG电泳图谱中的分布规律,并对一些特殊的电泳和分子杂交现象进行了讨论。     

GENETIC LOAD OF THE YEAST SACCHAROMYCES CEREVISIAE UNDER DIVERSE ENVIRONMENTAL CONDITIONS

Fitness effect of spontaneous mutations accumulated in mismatch-repair deficient strains of yeast was estimated by measuring their maximum growth rate. Several environments with different energetic substrates, nutritional conditions, and temperature were tested. Genetic load of haploid strains was about 20–30% under most of these conditions. Because such a pronounced effect was caused by relatively small lesions (point mutations) affecting probably less than 1% of genes, resistance of the yeast genome to DNA damage appears to be rather limited. Fitness transitions among environments were orderly, in the sense that some strains tended to be more or less fit than others in all circumstances. One of the environments (an extremely high temperature, 38°C) was stressful to the strains that accumulated mutations, as some of them stopped to grow, whereas the mutation-free strains were only moderately affected. These results imply that the impact of random point mutations is substantial and generally not dependent on a particular environment. Under stressful conditions, however, natural selection may be especially effective in purging mutations that, if commonly met, could slow down the rate of mutation accumulation.     

丙酮酸野生酵母菌的筛选及其生理生化特性研究

利用各种方法从来自苏锡地区的多种样品中广泛分离出200种野生酵母,然后借助纸上层析,获得13株具有纯丙酮酸生产能力的菌株.对其中2株的生理生化特性展开了进一步研究,初步认为其中1株性状类似南极洲假丝酵母(Candida antarctica),另1株属光滑球拟酵母(Torulopsis glabrata).