拮抗菌株Kc-t99的鉴定及其抑菌活性研究

通过传统分类与分子生物学技术相结合的方法对从京郊菜园土壤中分离筛选的拮抗菌株Kc-t99进行鉴定,并采用生物测定的方法评价其抑菌活性.结果表明,菌株Kc-t99的形态学特征和生理生化特性与枯草芽孢杆菌基本一致,其16S rDNA序列与GenBank中已鉴定的枯草芽孢杆菌的16S rDNA序列同源性高达98.06%,据此初步确定其为枯草芽孢杆菌.抑菌试验表明该菌株对供试的5种病原真菌和4种细菌均具抑菌活性,其中对甘蓝枯萎病菌、黄瓜角斑病菌和桃褐腐病菌抑制作用显著.     

细菌群体感应抑制活性微生物Zou 03的筛选与鉴定

从近海区生态环境中分离纯化98株海洋菌株,以根癌农杆菌WCF47为敏感检测菌株,筛选出1株具细菌群体感应抑制活性的菌株Zou03,对其进行形态、生理生化特征鉴定和16S rDNA分子鉴定。结果显示,Zou03具枯草芽胞杆菌(Bacillus subtilis)的典型特征,其16S rDNA序列通过对比分析,与GenBank中枯草芽胞杆菌16SrDNA的部分序列同源性为100%。综合形态、生化特征及16S rDNA序列对比分分析,鉴定菌株Zou03为枯草芽胞杆菌。表明近海区生态环境中存在具有抑制细菌群体感应活性的微生物,有利于海洋微生物资源开发,为以致病菌群体感应系统为靶点的新型疗法提供新技术。     

一株引起马来甜龙竹组培污染内生菌的分离与鉴定

【目的】对一株引起马来甜龙竹组培污染内生菌的分离与鉴定。【方法】采用改良的NA培养基分离纯化菌株,并通过菌体的形态结构观察、生理生化试验及其16SrDNA序列同源性分析对其进行鉴定。【结果】菌株SWFU01的形态特征及生理生化试验结果与解淀粉芽孢杆菌[Bacillus amyloliquefaciens(Fukumoto)Priest et al.]的描述基本相同;16S rDNA序列分析表明,该菌株与解淀粉芽孢杆菌JS在同一系统发育分支,其同源性为99.28%。【结论】综合形态学特征、生理生化特征以及16S rDNA序列分析的研究结果,菌株SWFU01被鉴定为解淀粉芽孢杆菌。     

一株细薄星芒海绵细菌的抗菌活性与分类学研究

采用平板涂布法从我国南海细薄星芒海绵分离得到一株细菌A72,以大肠埃希氏菌、金黄色葡萄球菌、枯草芽孢杆菌、荧光假单胞菌、黑曲霉、白假丝酵母、宛氏拟青霉7种指标菌对A72的抗菌活性进行了研究,同时采用形态学观察、生理生化分析与16S rDNA同源性与系统发育分析进行种属鉴定。研究发现A72对于枯草芽孢杆菌等具有显著的活性,初步确认A72为粪产碱杆菌。     

四株抗荔枝病原菌的芽孢杆菌的分离鉴定

荔枝果树根际土壤中筛选出4株芽孢杆菌OR-1、OR-2、OR-3、ON-6,都显示出抗荔枝病原菌的活性。采取对该菌株形态特征、培养特征、生理生化特征和遗传特性进行研究的方法,结果表明菌株与枯草芽孢杆菌(Bacillus subtilis)的特征一致;将4菌株的16S rDNA序列在GenBank中进行序列比对,结果亦显示其与Bacillus subtilis的16S rDNA的序列片段的相似性均达99%以上;以相似性为基础构建系统进化树,分析表明菌株与Bacillus subtilis同源关系最近。最终得出结论为菌株OR-1、OR-2、OR-3、ON-6为枯草芽孢杆菌。     

一株多功能芽孢杆菌的鉴定

【摘 要】 目的 从土壤筛选所得1株芽孢杆菌并对该菌进行鉴定.方法 通过表型特征、生理生化特性和16S rDNA序列同源性分析（Gen Bank登录号为: JN609382）。结果 形态学观察：菌落呈圆形,白色微黄,中央凸起,半透明,长时间培养容易形成褶皱;革兰染色为阳性;芽孢中生,周生鞭毛,细长且多。生理生化特征：参照《常见细菌手册》和《伯杰氏细菌鉴定手册》,其中接触酶反应、V-P测定、卵磷脂以及革兰染色呈阳性;能够利用D-葡萄糖、D-木糖、D-甘露醇产酸,并能使淀粉水解;不能利用丙酸盐,不能形成吲哚,不能水解酪氨酸,生理生化试验结果显示该芽孢杆菌与枯草芽孢杆菌的特征一致。分子生物学鉴定：在Gen Bank上经过BLAT分析,其与枯草芽孢杆菌的相似性最高（99％）。结论 鉴定该菌为枯草芽孢杆菌。     

产广谱细菌素乳酸菌的筛选和鉴定

以大肠杆菌、金黄色葡萄球菌、藤黄微球菌、铜绿假单胞杆菌和枯草芽孢杆菌为指示菌,从分离自四川传统发酵食品中的267株乳酸菌中,采用平板打孔法初筛、牛津杯法复筛(排除酸、过氧化氢干扰以及胰蛋白酶和木瓜蛋白酶处理),筛选出1株分离自醪糟的具有较强抑菌作用的产广谱细菌素的乳杆菌菌株P158,结合形态学、生理生化特性和16S rDNA序列同源性分析,该菌株被鉴定为植物乳杆菌(Lactobacillus plantarum)。     

菌株F12-11-1-2的16S rDNA序列分析及其生理生化性质研究

菌株F12-11-1-2是一株从中国东海浙江海域200m的海泥中分离得到的,具有抗稻瘟霉(Pyricularia oryzae)活性。通过对菌株的形态、培养特征、生理生化特征的研究以及16S rDNA序列分析,结果表明它是一株适应了海洋环境的芽孢杆菌属(Bacillus)的枯草芽孢杆菌(Bacillus subtilis)。     

三株芸豆叶际拮抗细菌的筛选与鉴定

以芸豆锈病病原菌[Uromyces appendiculatus(Pers.)Ung.]为指示菌,从芸豆叶际中筛选到3株具有明显拮抗效果的细菌,编号为SS2,L14b,NEW2。经过形态学观察,生理生化测定,16S rDNA序列及系统发育分析,初步鉴定SS2为短短芽孢杆菌(Brevibacillus brevis),L14b与NEW2为枯草芽孢杆菌(Bacillus subtilis)。SS2,L14b,NEW2的16S rDNA序列的Gen-Bank登录号分别为EU771078、EU771076和EU771079。     

产广谱细菌素乳酸菌的筛选和鉴定

以大肠杆菌、金黄色葡萄球菌、藤黄微球菌、铜绿假单胞杆菌和枯草芽孢杆菌为指示菌,从分离自四川传统发酵食品中的267株乳酸菌中,采用平板打孔法初筛、牛津杯法复筛（排除酸、过氧化氢干扰以及胰蛋白酶和木瓜蛋白酶处理）,筛选出1株分离自醪糟的具有较强抑菌作用的产广谱细菌素的乳杆菌菌株P158,结合形态学、生理生化特性和16S rDNA序列同源性分析,该菌株被鉴定为植物乳杆菌（Lactobacillus plantarum）。     

Depth-dependent strain of patellofemoral articular cartilage in unconfined compression

This biomechanical study reports strain gradients in patellofemoral joint cross-sections of seven porcine specimens in response to 1% unconfined axial compression subsequent to specific amounts of off-set strain. Strain distributions were quantified with a customized laser-based electronic speckle pattern interferometry (ESPI) system in a non-contact manner, delivering high-resolution, high-sensitivity strain maps over entire patellofemoral cartilage cross-sections. Strain reports were evaluated to determine differences in strain magnitudes between the superficial, middle, and deep cartilage layers in femoral and patellar cartilage. In addition, the effect of 5%, 10%, 15%, and 20% off-set strain on depth-dependent strain gradients was quantified. Regardless of the amount of off-set strain, the superficial layer of femoral cartilage absorbed the most strain, and the deep layer absorbed the least strain. These depth-dependent strain gradients were most pronounced for 5% off-set strain, at which the superficial layer absorbed on average 5.7 and 23.7 times more strain as compared to the middle and deep layers, respectively. For increased off-set strain levels, strain gradients became less pronounced. At 20% off-set strain, differences in layer-specific strain were not statistically significant, with the superficial layer showing a 1.4 fold higher strain as the deep layer. Patellar cartilage exhibited similar strain gradients and effects of off-set strain, although the patellar strain was on average 19% larger as compared to corresponding femoral strain reports. This study quantified for the first time continuous strain gradients over patellofemoral cartilage cross-sections. Next to provision of a detailed functional characterization of normal diarthrodial joints, this novel experimental approach holds considerable attraction to investigate joint degenerative processes.     

Network Relationships of Bacteria in a Stable Mixed Culture

We investigated the network relationships of bacteria in a structurally stable mixed culture degrading cellulose. The mixed culture consists of four bacterial strains (a cellulose-degrading anaerobe [strain S], a saccharide-utilizing anaerobe [strain F], a peptide- and acetate-utilizing aerobe [strain 3] and a peptide-, glucose-, and ethanol-utilizing aerobe [strain 5]). Interspecies interactions were examined by analyzing the effects of culture filtrates on the growth of the other strains and by comprehensively analyzing population dynamics in the mixed-culture systems with all possible combinations of the four bacterial strains. The persistence of strain S depends on the effects of strain 5. However, strain 5 is a disadvantaged strain because strain 3 has bacteriocidal activity on strain 5. The extinction of strain 5 is indirectly prevented by strain F that suppresses the growth of strain 3. Although strain F directly has suppressive effects on the growth of strain S, strain F is essential for the persistence of strain S, considering the indirect effects (maintaining strain 5, which is essential for the survival of strain S, by inhibiting strain 3). These indirect relationships form a bacterial network in which all the relationships including suppressive effects were well balanced to maintain the structural stability. In addition to direct metabolite interactions, such kind of indirect relationships could have a great impact on microbial community structure in the natural environment.     

黑曲霉原生质体诱变选育β-葡萄糖苷酶高产菌株

本研究报道了以原生质体诱变技术选育高产β-葡萄糖苷酶的黑曲霉菌株,并研究了其发酵特性。以黑曲霉CGMCC3.316为出发菌株,通过紫外诱变得到突变株3-3M。然后以3-3M为供试菌株,研究了其原生质体制备与再生的条件。最后通过原生质体诱变,选育得到一株β-葡萄糖苷酶活力较高的突变株60B-3D。该菌株具有良好的遗传稳定性,酶活力平均达到23IU/mL,与出发菌株CGMCC3.316相比提高39%。此外,该菌株的木聚糖酶活力也有所增加。同时考察了黑曲霉60B-3D的发酵特性,并与3-3M和出发菌株进行比较,结果表明该菌株有较高的蛋白分泌能力。本研究为发酵生产β-葡萄糖苷酶提供了一株良好的供试菌株。     

Transductional construction of a threonine-producing strain of Serratia marcescens.

A threonine-producing strain of Serratia marcescens Sr41 was constructed according to the following process. Thr- strain E-60 was derived from strain HNr59 having constitutive levels of threonine-sensitive aspartokinase and homoserine dehydrogenase. Thr+ transductant T-570 was constructed from strain E-60 and phage grown on strain HNr21 having feedback-resistant threonine-sensitive aspartokinase and homoserine dehydrogenase. This transductant lacked both feedback inhibition and repression for the two enzymes. Thr- strain N-11 was derived from strain AECr174 lacking feedback inhibition and repression of lysine-sensitive aspartokinase. Subsequently, the threonine region of strain T-570 was transduced into strain N-11. One of the THR+ transductants, strain T-693, produced markedly high levels of the two aspartokinases and homoserine dehydrogenase, which were insensitive to feedback inhibition. This strain produced about 25 mg of threonine per ml in the medium containing sucrose and urea.     

Cloning and Characterization of the Genomic RNA Sequence of the Mumps Virus Strain Associated with a High Incidence of Aseptic Meningitis

cDNA clones of the mumps virus wild-type strain, associated with a high incidence of aseptic meningitis (ODATE-1 strain), were isolated and analyzed from genomic nucleotide position 22 to 8520 containing the NP, P, M., F, SH and HN protein coding region. The ODATE-1 strain exhibited a RFLP profile identical to that of the Urabe vaccine strain in spite of the fact that the virus was isolated from non-vaccinated cases. However, a comparison of nucleotide and amino acid sequences among the ODATE-1 strain, Urabe strain and Miyahara strain revealed that the ODATE-1 strain was not related to the Urabe strain.     

Transductional construction of a threonine-producing strain of Serratia marcescens.

A threonine-producing strain of Serratia marcescens Sr41 was constructed according to the following process. Thr- strain E-60 was derived from strain HNr59 having constitutive levels of threonine-sensitive aspartokinase and homoserine dehydrogenase. Thr+ transductant T-570 was constructed from strain E-60 and phage grown on strain HNr21 having feedback-resistant threonine-sensitive aspartokinase and homoserine dehydrogenase. This transductant lacked both feedback inhibition and repression for the two enzymes. Thr- strain N-11 was derived from strain AECr174 lacking feedback inhibition and repression of lysine-sensitive aspartokinase. Subsequently, the threonine region of strain T-570 was transduced into strain N-11. One of the THR+ transductants, strain T-693, produced markedly high levels of the two aspartokinases and homoserine dehydrogenase, which were insensitive to feedback inhibition. This strain produced about 25 mg of threonine per ml in the medium containing sucrose and urea.     

Alternation of the Dissimilation Pathway for Glycerol and Amplification of Glycerol Dehydrogenase in Mutant Strains of Cellulomonas sp. NT3060

Mutant strain ME544, which is able to grow on glycerol slowly, was derived from glycerol-negative mutant strain G011, which is a derivative strain of Cellulomonas sp. NT3060 and is defective in both the enzyme activities of glycerol kinase and glycerol 3-phosphate dehydrogenase. The mutant strain still lacked both the enzyme activities involved in the dissimilation of glycerol and had the same level of glycerol dehydrogenase activity as the parent strain. Dihydroxyacetone kinase activity in mutant strain ME544 was inducibly formed, reaching 4-fold the level in mutant strain G011 in glycerol medium. Thus, the mutant strain seemed to dissimilate glycerol by means of glycerol dehydrogenase followed by an increase in dihydroxyacetone kinase. Subsequently, a mutant strain, GP1807, which was resistant to the inhibition of growth on glycerol by 1,2-propanediol, was derived from mutant strain ME544. Glycerol dehydrogenase activity of the mutant strain was amplified about 6-fold compared to that of the wild type strain.     

The effect of strain rate on the mechanical properties of human cortical bone

Bone mechanical properties are typically evaluated at relatively low strain rates. However, the strain rate related to traumatic failure is likely to be orders of magnitude higher and this higher strain rate is likely to affect the mechanical properties. Previous work reporting on the effect of strain rate on the mechanical properties of bone predominantly used nonhuman bone. In the work reported here, the effect of strain rate on the tensile and compressive properties of human bone was investigated. Human femoral cortical bone was tested longitudinally at strain rates ranging between 0.14-29.1 s(-1) in compression and 0.08-17 s(-1) in tension. Young's modulus generally increased, across this strain rate range, for both tension and compression. Strength and strain (at maximum load) increased slightly in compression and decreased (for strain rates beyond 1 s(-1)) in tension. Stress and strain at yield decreased (for strain rates beyond 1 s(-1)) for both tension and compression. In general, there seemed to be a relatively simple linear relationship between yield properties and strain rate, but the relationships between postyield properties and strain rate were more complicated and indicated that strain rate has a stronger effect on postyield deformation than on initiation of yielding. The behavior seen in compression is broadly in agreement with past literature, while the behavior observed in tension may be explained by a ductile to brittle transition of bone at moderate to high strain rates.     

格特隐球菌HOG1基因的敲除及重建

目的 构建格特隐球菌HOG1基因缺陷株和HOG1基因重建株.方法 从格特隐球菌基因组扩增HOG1基因,通过部分基因缺失方法,获得缺陷基因dHOG1.将获得的HOG1基因及其缺陷基因dHOG1分别亚克隆到真核表达载体pGAPzα-A,构建pGAPzα-HOG1及pGAPzcα-dHOG1质粒.将pGAPzα-dHOG1质粒转染隐球菌原始株,通过筛选获得HOG1基因缺陷菌株;同样方法将pGAPzα-HOG1质粒转染格特隐球菌HOGI基因敲除菌株,获得HOGI基因重建株.结果 RTPCR结果示:格特隐球菌HOG1基因缺陷株不转录表达完整的HOG1基因,而HOG1基因重建株可以转录表达完整的HOG1基因片段.结论 成功获得格特隐球菌HOG1缺陷株和HOG1基因重建株,为后续格特隐球菌毒力和致病机制的研究奠定了基础.     

phbC基因敲除对土壤杆菌产可得然胶的影响

本文构建了phbC基因无痕敲除菌株ΔphbC,分析了ΔphbC菌株生长代谢情况和产生的可得然胶在产量、凝胶性质和红外结构的变化。结果显示,ΔphbC菌株在发酵过程中氨基氮消耗情况与野生型菌株一致,在蔗糖消耗方面,ΔphbC菌株与野生型菌株在18 h之后出现显著差异,蔗糖消耗比野生型菌株明显降低。ΔphbC菌株可得然胶产量约24 g/L,相对于野生型菌株降低了45%;胶凝胶强度为812.521 g/cm^2,相对于野生型菌株降低了21%;红外结构与野生型菌株一致,无明显差异。phbC基因不影响菌体生长,不影响可得然胶结构,但是影响可得然胶的合成。