乙酰乳酸合成酶的制备、分离与纯化

乙酰乳酸合成酶［１］（ａｃｅｔｏｌａｃｔａｔｅｓｙｎｔｈａｓｅ,ＡＬＳ）是目前国际上除草剂研究的前沿热点．它能催化一分子丙酮酸脱羧并与另一分子丙酮酸缩合成一分子乙酰乳酸,从而导致侧链氨基酸缬氨酸、亮氨酸和异亮氨酸的生物合成．存在于细菌和植物组织中［２...     

中华鳖白底板病和红底板病细菌的分离鉴定及致病性

从患白底板病和红底板病的病鳖中分离出７株细菌,应用一般细菌分离鉴定的方法和美国生物－梅里埃（Ｂｉｏ Ｍｅｒｉｅｕｘ ＵＳＡ）公司ＶＩＴＥＫ全自动微生物鉴定系统进行鉴定,结果显示有３株嗜水气单胞菌（Ａｅｒｏｍｏｎａｓ ｈｙｄｒｏｐｈｉｌａ）、１株温和气单胞菌（Ａｅｒｏｍｏｎａｓ ｓｏｂｒｉａ<.I>）、１株金黄色葡萄球菌（Ｓｔａｐｈｙｌｏｃｏｃｃｕｓａｕｒｅｕｓ）、１株肺炎克雷伯氏菌（Ｋｌｅｂｓｉｅｌｌａ ｐｎｅｕｍｏｎｉａｅ）和 １株美人鱼弧菌（Ｖｉｂ     

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文摘基因工程９８００９８血清中小型基因组ＤＮＡ片段的直接ＰＣＲ［英］／Ｓａｎｄｆｏｌｄ,Ａ．Ｊ．…∥ＢｉｏＴｅｃｈｎｉｑｕｅｓ．－１９９７,２３（５）．－８９０～８９２［译自ＤＢＡ,１９９８,１７（１）,９８－０００１９］开发了一种简易、快速的方法,...     

［B1－Ala，B2－Ala］－胰岛素：受体结合特性及生物活力研究

本文报道了［Ｂ１～Ａｌａ,Ｂ２－Ａｌａ］－胰岛素与人胎盘细胞膜胰岛素受体结合的特性和体外生物活力,并与胰岛素进行比较。在３７℃和杆菌肽存在下,１２５Ｉ－［Ｂ１－Ａｌａ,Ｂ２－Ａｌａ］－胰岛素和１２５Ｉ－胰岛素与人胎盘细胞膜作用依赖于反应时间,反应６分钟到达平衡,此时,［Ｂ１－Ａｌａ,Ｂ２－Ａｌａ］－胰岛素和胰岛素与胰岛素受体的最大结合分别为每毫克膜蛋白结合６．４４ｆｍｏｌ和３．４７ｆｍｏｌ：达到平衡一半所需时间（Ｔ１／２）分别为１９秒和２５秒。用１２５Ｉ－［Ｂ１－Ａｌａ,Ｂ２－Ａｌａ］－胰岛素作为放射配体进行竞争性结合研究,从ＩＣ（５０）得［Ｂ１－Ａｌａ,Ｂ２－Ａｌａ］胰岛素的受体结合活力为胰岛素的１３９．６％。Ｓｃａｔｏｈａｒｄ分析求得;［Ｂ１－Ａｌａ,Ｂ２－Ａｌａ］－胰岛素与高亲和和低亲和结合位点的结合常数在４℃时分别为５．８８×１０８Ｌ／ｍｏｌ和７．６３×１０５Ｌ／ｍｏｌ,而胰岛素分别为４．８３×１０８Ｌ／ｍｏｌ和３．３９×１０５Ｌ／ｍｏｌ。促脂肪细胞生成脂的实验表明：［Ｂ１－Ａｌａ,Ｂ２－Ａｌａ］－胰岛素的活力为胰岛素的１３０％。     

瓜叶乌头的二萜生物碱

从瓜叶乌头（ＡｃｏｎｉｔｕｍｈｅｍｓｌｅｙａｎｕｍＰｒｉｔｚ．）的块根分离到４个二萜生物碱,通过波谱分析和化学方法分别鉴定为８－ａｃｅｔｙｌ－１４－ｂｅｎｚｏｙｌ－ｅｚｏｃｈａｓｍａｎｉｎｅ（１）、ｃｈａｓｍａｎｉｎｅ（２）、ｉｎｄａｃｏｎｉｔｉｎｅ（３）和ｔａｌａｔｉｓａｍｉｎｅ（４）。其中１为新化合物,命名为瓜叶乌宁（ｈｅｍｓｌｅｙａｎｉｎｅ）。瓜叶乌宁（１）为白色块状结晶,ｍｐ１１５—１１６℃,［α］２０Ｄ＋１９．０２°。由ＦＡＢ－ＭＳ和ＥＩ－ＭＳ得知,分子量为６１３,１３Ｃ－ＮＭＲ中３２…     

草玉梅化学成分的研究

草玉梅（ＡｎｅｍｏｎｅｒｉｖｕｌａｒｉｓＢｕｃｈ．－Ｈａｍ．ｅｘＤＣ．）为毛茛科银莲花属植物,根状茎和叶供药用,治疗喉炎、扁桃腺炎、风湿疼痛和胃痛等症［１］。其化学成分已有报道［２,３］,主要为三萜皂甙。最近,我们从草玉梅全草的乙醇提取物中分离鉴定了３种化学成分,其中两种为已知成分乌苏酸（Ⅰ）和β－谷甾醇（Ⅱ）,一种为新化合物,命名为草玉梅甙Ａ（ａｎｅｍｏｓｉｄｅＡ）（Ⅲ）。本文主要报道草玉梅甙Ａ的结构鉴定。化合物Ⅲ为白色粉末,Ｌｉｅｂｅｒｍａｎｎ－Ｂｕｒｃｈａｒｄ试验为阳性,Ⅲ经酸水解,甙元与…     

六倍体山羊草与普通小麦杂种F1细胞学研究

粗厚山羊草［Ａｅｇｉｌｏｐｓｃｒａｓｓａ Ｂｏｉｓｓ．（２ｎ＝ ６ｘ＝ ４２,ＤＤＤ２ Ｄ２ＭＣｒＭＣｒ）］、叙利亚山羊草［Ａｅ．ｖａｖｉｌｏｖｉｉ （Ｚｈｕｋ．）Ｃｈｅｎｎ．（２ｎ＝ ６ｘ＝ ４２, ＤＤ ＭＣｒＭＣｒＳＰ ＳＰ）］与普通小麦［Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．（２ｎ＝ ６ｘ＝ ４２, ＡＡＢＢＤＤ）］杂种Ｆ１ 植株形态大多偏向山羊草亲本。４ 个叙利亚山羊草×普通小麦和１ 个粗厚山羊草×普通小麦杂种Ｆ１ 自交结实,结实率在０．１％ —６．５％ 之间。这些种子胚乳很少,生活力较弱,播种后,只有少数种子出苗。杂种Ｆ１ ＰＭＣ减数分裂染色体配对水平较低,出现大量的单价体,二价体低于理论值,并且大多为棒形,说明两种山羊草的Ｄ组染色体已经过很大的修饰。在各杂种Ｆ１ 中还观察到少量的三价体,有些杂种还可见频率很低的四价体和五价体。以山羊草为母本的杂种Ｆ１ 染色体交叉频率优于反交。Ｆ１ 染色体分离极不正常,产生大量的多分孢子和微核。在叙利亚山羊草×冀麦３０ 号中还发现１ 株体细胞染色体为２１ 条的植株,其原因尚需进一步研究确定     

蜘蛛抱蛋根茎中的甾体皂甙

从蜘蛛抱蛋（ＡｓｐｉｄｉｓｔｒａｅｌａｔｉｏｒＢｌ．）根茎的甲醇提取物中分离得两个甾体皂甙。经波谱解析及化学降解证明其化学结构分别为薯芋皂甙元一３一Ｏ－［Ｂ－Ｄ－葡萄吡喃糖基（１→２）］－［Ｂ－Ｄ－木吡喃糖基（１→３）］－Ｂ－Ｄ－葡萄吡喃糖基（１→４）－Ｂ－Ｄ一半乳吡喃糖甙（即蜘蛛抱蛋甙ａｓｐｉｄｉｓｔｒｉｎ）及新静诺特皂甙元－３－Ｏ－［Ｂ－Ｄ－葡萄吡喃糖基（１→２）］～［Ｂ－Ｄ－木吡喃糖基（１→３）－Ｂ－Ｄ－葡萄吡喃糖基（１→４）一Ｂ－Ｄ－半乳吡喃糖甙,后者为一新试,命名为新蜘蛛抱蛋甙ｒｅｏａｓｐｉｄｉｓｔｒｉｎ。     

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文摘基因工程９８０００１用荧光假单胞菌不同突变株接种的土壤中的微生物活性［英］／Ｋｏｚｄｒｏｊ,Ｊ．∥ＡｃｔａＭｉｃｒｏｂｉｏｌ．Ｐｏｌ．－１９９７,４６（１）．－５７～６３［译自ＤＢＡ,１９９７,１６（１８）,９７－０９３０９］研究了荧光假单胞菌...     

虎刺楤木根皮化学成分研究

从虎刺楤木［Ａｒａｌｉａ ａｒｍ ａｔａ （Ｗａｌｌ．） Ｓｅｅｍ ．］根皮中分离到１１个成分,根据理化及光谱性质,鉴定其中７个为皂甙,即去葡萄糖竹节参皂甙Ⅳａ（ＡＴ－Ⅰ）、竹节参皂甙Ⅳａ（ＡＴ－Ⅱ）、姜状三七甙Ｒ１（ＡＴ－Ⅲ）、人参皂甙Ｒ０（ＡＴ－Ⅳ）、Ａｄ－Ⅲ［（ＡＴ－Ⅴ）后称黄毛楤木皂甙］、虎刺楤木皂甙（ＡＴ－Ⅵ）和楤木皂甙Ａ（ＡＴ－Ⅶ）,其余４个成分则为二十八羧酸（ＡＴ－Ⅷ）、谷甾醇（ＡＴ－Ⅸ）、谷甾醇与豆甾醇的混合物（ＡＴ－Ⅹ）及齐墩果酸（ＡＴ－Ⅺ）。以上成分均系首次从该植物中分得,其中ＡＴ－Ⅵ则为一新皂甙     

Survival, mutation and capacity to repair single-strand DNA breaks after gamma irradiation in different Exr? strains of Escherichia coli

Summary Strains of Escherichia coli K-12 and B/r made by transduction of the exrA allele from a B_s-2 derivative have been compared with Exr(W)^– strains derived from B_s-1 and B_s-2 by mutation (E.M. Witkin, 1967). Both transduced exrA and Exr(W)^– strains were almost unmutable by gamma radiation, but the former class were as sensitive to gamma radiation as recA strains and, like them, were unable to repair single-strand DNA breaks as detected by the McGrath-Williams technique. In contrast the Exr(W)^–strains were as resistant to gammaradiation as Exr(W)⁺ strains derived from them and were equally efficient in repairing single-strand breaks. The existence of Exr(W)^–strains suggests that the mutagenicity of single-strand breaks may depend entirely on the way in which they are repaired. The properties of the (Exr(W)^–strains cannot be ascribed solely to the transducable exrA allele.A large effect of diffuse daylight in lowering the molecular weight of DNA on alkaline sucrose gradients is described which, unless prevented, may lead to erroneous results in such experiments.     

甘蔗赤腐病内生拮抗菌YC89的筛选及鉴定

【目的】筛选防治甘蔗赤腐病(sugarcane red rot)的生防菌株。【方法】实验以前期分离获得的甘蔗内生细菌为目标菌,以甘蔗赤腐病的病原真菌镰孢炭疽菌(Colletotrichum falcatum Went.)为指示菌,采用平板对峙法筛选对该病菌有较强抑制作用的菌株,然后通过琼脂扩散法测定菌株代谢产物对抑菌活性的影响,并对具有较好拮抗效果的高效菌株进行抑菌广谱性分析并对其进行鉴定。最后通过形态学、生理生化特征以及16SrDNA和gyrA序列分析对高效菌株YC89进行鉴定。【结果】经初筛筛选到抑菌带均大于1.60 cm的5株拮抗细菌,其中X22、W2、YC89抑菌带均高达1.87 cm。对初筛得到的5株内生菌进行复筛,结果所示菌株YC89、H1、X22、W2、YT93对镰孢炭疽菌的抑菌率都在75%以上,其中菌株YC89对该病菌的抑菌效果最好,其抑菌率为78%。菌株YC89的发酵液、上清液、过滤液及粗蛋白提取液对镰孢炭疽菌的生长有较强的抑制作用,且菌株YC89对玉米大斑病、甘蔗梢腐病、草莓灰霉病等7种病原菌也有较好的抑制效果。通过菌株鉴定结果,初步将YC89菌株鉴定为贝莱斯芽孢杆菌(Bacillus velezensis)。【结论】菌株YC89对镰孢炭疽菌具有较好的抑制效果,表明其对甘蔗赤腐病的生物防治具有较大的应用潜力。     

Isolation and characterization of fructophilic lactic acid bacteria from fructose-rich niches

Fourteen strains of fructophilic lactic acid bacteria were isolated from fructose-rich niches, flowers, and fruits. Phylogenetic analysis and BLAST analysis of 16S rDNA sequences identified six strains as Lactobacillus kunkeei, four as Fructobacillus pseudoficulneus, and one as Fructobacillus fructosus. The remaining three strains grouped within the Lactobacillus buchneri phylogenetic subcluster, but shared low sequence similarities to other known Lactobacillus spp. The fructophilic strains fermented only a few carbohydrates and fermented d-fructose faster than d-glucose. Based on the growth characteristics, the 14 isolates were divided into two groups. Strains in the first group containing L. kunkeei, F. fructosus, and F. pseudoficulneus grew well on d-fructose and on d-glucose with pyruvate or oxygen as external electron acceptors, but poorly on d-glucose without the electron acceptors. Strains in this group were classified as “obligately” fructophilic lactic acid bacteria. The second group contained three unidentified strains of Lactobacillus that grew well on d-fructose and on d-glucose with the electron acceptors. These strains grew on d-glucose without the electron acceptors, but at a delayed rate. Strains in this group were classified as facultatively fructophilic lactic acid bacteria. All fructophilic isolates were heterofermentative lactic acid bacteria, but “obligately” fructophilic lactic acid bacteria mainly produced lactic acid and acetic acid and very little ethanol from d-glucose. Facultatively fructophilic strains produced lactic acid, acetic acid and ethanol, but at a ratio different from that recorded for heterofermentative lactic acid bacteria. These unique characteristics may have been obtained through adaptation to the habitat.     

Antibiotic properties of the strains of the basidiomycete Lentinus edodes (Berk.) sing]

Antibiotic properties of the extracts from the fermentation broth and mycelium of 15 strains of the edible and medicinal basidiomycete L. edodes were studied and it was shown that the extracts were active against grampositive and gramnegative bacteria, yeasts and mycelial fungi, including dermatophytes and phytopathogens. The strains differed by the set of the organisms susceptible to the action of the extracts. Strains of L. edodes combining marked antibiotic properties and high yields of water soluble polysaccharides were screened. The active compounds were detected by preparative TLC. Two of them were identified with UV- and mass spectrometry as lentinamycin B and erytadenine (lentinacin). Lentinamycin B was found to be the main component responsible for the antibiotic activity of the L. edodes strains.     

High effectiveness of the method with cefpirome in detection of extended-spectrum beta-lactamases in different species of gram-negative bacilli

Gram-negative bacilli were examined for ESBL production by using four methods: double-disc synergy diffusion test (DDST), and three tests of combined discs with cefpodoxime, ceftazidime and cefotaxime alone and the same cephalosporins with clavulanic acid. Strains determined as ESBL-negative with all these tests were examined by using fifth method with cefpirome. 47,5% from 178 negative in other methods strains, appeared ESBL-positive in this test. The examined strains belonged to 16 different species. Most of them were Enterobacter cloaceae, Serratia marcescens, Pseudomonas aeruginosa and Acinetobacter baumanii. It seems that the combined discs method with cefpirome may be usefull for phenotypic detection of ESBL producing bacteria also in the case of strains where ESBL production is camouflaged with derepressed chromosomal AmpC beta-lactamases.     

Screening of species-specific lactic acid bacteria for veal calves multi-strain probiotic adjuncts

The selection of promising specific species of lactic acid bacteria with potential probiotic characteristics is of particular interest in producing multi species-specific probiotic adjuncts in veal calves rearing. The aim of the present work was to select and evaluate in?vitro the functional activity of lactic acid bacteria, Bifidobacterium longum and Bacillus coagulans strains isolated from veal calves in order to assess their potential use as multi species-specific probiotics for veal calves. For this purpose, bacterial strains isolated from faeces collected from 40 healthy 50-day-calves, were identified by RiboPrinter and 16s rRNA gene sequence. The most frequent strains belonged to the species B. longum, Streptococcus bovis, Lactobacillus animalis and Streptococcus macedonicus. Among these, 7 strains were chosen for testing their probiotic characteristics in?vitro. Three strains, namely L. animalis SB310, Lactobacillus paracasei subsp. paracasei SB137 and B. coagulans SB117 showed varying individual but promising capabilities to survive in the gastrointestinal tract, to adhere, to produce antimicrobial compounds. These three selected species-specific bacteria demonstrated in?vitro, both singularly and mixed, the functional properties needed for their use as potential probiotics in veal calves.     

Historical microbiology: revival and phylogenetic analysis of the luminous bacterial cultures of M. W. Beijerinck

Luminous bacteria isolated by Martinus W. Beijerinck were sealed in glass ampoules in 1924 and 1925 and stored under the names Photobacterium phosphoreum and 'Photobacterium splendidum'. To determine if the stored cultures were viable and to assess their evolutionary relationship with currently recognized bacteria, portions of the ampoule contents were inoculated into culture medium. Growth and luminescence were evident after 13 days of incubation, indicating the presence of viable cells after more than 80 years of storage. The Beijerinck strains are apparently the oldest bacterial cultures to be revived from storage. Multi-locus sequence analysis, based on the 16S rRNA, gapA, gyrB, pyrH, recA, luxA, and luxB genes, revealed that the Beijerinck strains are distant from the type strains of P. phosphoreum, ATCC 11040(T), and Vibrio splendidus, ATCC 33125(T), and instead form an evolutionarily distinct clade of Vibrio. Newly isolated strains from coastal seawater in Norway, France, Uruguay, Mexico, and Japan grouped with the Beijerinck strains, indicating a global distribution for this new clade, designated as the beijerinckii clade. Strains of the beijerinckii clade exhibited little sequence variation for the seven genes and approximately 6300 nucleotides examined despite the geographic distances and the more than 80 years separating their isolation. Gram-negative bacteria therefore can survive for many decades in liquid storage, and in nature, they do not necessarily diverge rapidly over time.     

Adherence to HEp-2 cells and replication in macrophages of Salmonella derby of human origin.

Adherence to a HEp-2 cell monolayer was tested for in four strains of Salmonella derby which were isolated from patients with diarrhea. One strain, SB1, was highly adherent and another strain, SB4, was nonadherent. The other two strains exhibited moderate adherence. Further in vitro study of invasion of HEp-2 cells by S. derby and its replication in murine peritoneal macrophages was carried out using SB1 and SB4. Thin section electron micrographs revealed that SB1 invaded HEp-2 cells but SB4 did not. The number of viable bacteria within macrophages was determined at intervals after inoculation of bacteria. The result indicates that SB1 can replicate in the macrophages but SB4 cannot. Flagella and fimbriae were compared by electron microscopy between SB1 and SB4, and their lipopolysaccharides and outer membrane proteins were also compared with each other by SDS-polyacrylamide gel electrophoresis. The presence of a 41 kDa protein in the outer membranes of SB1 was only the difference detected, suggesting that this protein could be a factor required for adherence of this serovar to epithelial cells.     

Physiological and phylogenetic diversity of thermophilic spore-forming hydrocarbon-oxidizing bacteria from oil fields

The distribution and population density of aerobic hydrocarbon-oxidizing bacteria in the high-temperature oil fields of Western Siberia, Kazakhstan, and China were studied. Seven strains of aerobic thermophilic spore-forming bacteria were isolated from the oil fields and studied by microbiological and molecular biological methods. Based on the 16S rRNA gene sequences, phenotypic characteristics, and the results of DNA-DNA hybridization, the taxonomic affiliation of the isolates was tentatively established. The strains were assigned to the first and fifth subgroups of the genus Bacillus on the phylogenetic branch of the gram-positive bacteria. Strains B and 421 were classified as B. licheniformis. Strains X and U, located between B. stearothermophilus and B. thermocatenulatus on the phylogenetic tree, and strains K, Sam, and 34, related but not identical to B. thermodenitrificans and B. thermoleovorans, undoubtedly represent two new species. Phylogenetically and metabolically related representatives of thermophilic bacilli were found to occur in geographically distant oil fields.     

新疆棉花黄萎病菌拮抗细菌的分离、筛选与鉴定

【背景】棉花黄萎病是由大丽轮枝菌(Verticillium dahliae Kleb.)引起的一种世界性病害,近年来对该病害的生物防治因具有环境友好和人畜安全的特性而倍受关注。【目的】筛选棉花黄萎病高效拮抗细菌并对其进行鉴定,为棉花黄萎病的生物防治扩充菌种资源。【方法】采用稀释涂布平板法分离细菌,并进行拮抗细菌的初筛和复筛,通过形态特征、生理生化特征和16S rRNA基因序列分析对筛选到的细菌进行鉴定,确定其分类地位。【结果】初筛分离到535株对病原菌具有拮抗作用的细菌,并选取了108株拮抗细菌进行复筛,最终筛选到了4株优势拮抗细菌。通过形态观察、生理生化特征和16SrRNA基因序列分析,将菌株BHZ-29、SHT-15、SHZ-24和SMT-24分别鉴定为贝莱斯芽孢杆菌(Bacillusvelezensis)、枯草芽孢杆菌斯皮兹仁亚种(Bacillus subtilis subsp. spizizenii)、萎缩芽孢杆菌(Bacillus atrophaeus)和香草芽孢杆菌(Bacillus vanillea)。【结论】获得了4株高效拮抗细菌,并且首次报道了香草芽孢杆菌对棉花黄萎病菌具有抑制作用。