产聚β-羟基丁酸酯菌株的筛选及发酵条件的优化

作为一类可望替代传统塑料的新型可降解生物高分子材料,聚β-羟基丁酸酯(PHB)日益引起人们的重视。采用尼罗蓝荧光法从污水中初筛得到产PHB的细菌,摇瓶发酵复筛得到一株PHB产量较高的菌株AE13,同时对该菌株的发酵条件进行了正交优化,PHB的产量达到0.85g/L。     

紫外线诱变选育高产PHB解聚酶的菌株

以降解聚-β羟基丁酸酯(PHB)的青霉(Penieillium sp．)DS9713a为出发菌株,通过紫外线(UV)诱变分生孢子,采用透明圈初筛和摇瓶复筛,获得酶活高于原始菌株的突变株5株,其中DS9713a-CS01突变株的PHB解聚酶活力高于对照97．42％,并对其酶学性质进行了初步研究。     

合成聚β-羟基丁酸(PHB)鞘细菌的分离、鉴定与筛选

通过富集培养和划线分离等手段从活性污泥中分离筛选到一株合成PHB鞘细菌FQ4 0 ,其生物学特征为 :杆菌 ,大小 (0 .6～ 1.5 ) μm× (1.5～ 5 .5 ) μm ;革兰氏染色阴性 ;具衣鞘 ,丝状体很长 ,偶见假分枝 ;亚端生鞭毛 ;胞内具PHB颗粒 ;菌落为光滑型和丝状型两种。经鉴定该菌株为鞘细菌类球衣菌属中的浮游球衣菌 (Sphaerotilusnatans)。对其合成PHB研究表明 :细胞内可积累大量的PHB颗粒 ,含量可达细胞干重的 30 .5 %。     

紫外线诱变选育高产PHB解聚酶的菌株*

以降解聚-β羟基丁酸酯(PHB)的青霉(Penicilliumsp.)DS9713a为出发菌株,通过紫外线(UV)诱变分生孢子,采用透明圈初筛和摇瓶复筛,获得酶活高于原始菌株的突变株5株,其中DS9713a-CS01突变株的PHB解聚酶活力高于对照97.42%,并对其酶学性质进行了初步研究。     

一株新型高产PHB假单胞菌SCH17的分离和特征分析

通过丁醇富集筛选,从土壤样品中筛选到一株菌株SCH17。经过生理特性和16S rRNA分析,鉴定菌株SCH17属于假单胞菌属。透射电镜显示该菌细胞内聚集了大量颗粒状物质,经过氯仿抽提和核磁共振分析,确认这些颗粒物质是聚β-羟基丁酸(PHB)。通过对碳源和氮源的优化,得到最佳积累PHB的碳源是果糖,氮源是蛋白胨。在该培养基中仅需发酵14 h,菌体干重和PHB含量均达到最大,分别为3.52 g/L和2.69 g/L,PHB含量高达细胞干重的76%。     

漆酶高产菌株的筛选及产酶条件研究

木质素是一种高度复杂的不定形的芳香族化合物 ,是仅次于纤维素的第二大再生有机资源 ,木质素的微生物降解及其有关酶类在制浆造纸工业和环境保护方面具有较大潜力。木质素的生物降解酶主要有木素过氧化物酶、漆酶、锰过氧化物酶和多酚氧化酶等。国外已有该方面的研究报道[5～ 7,9,10 ] ,但多集中在黄孢原毛平革菌 (Ｐｈａｎｅｒｏｃｈａｅｔｅｃｈｒｙｓｏｓｐｏｒｉｕｍ)木素过氧化物酶和锰过氧化物酶方面的研究 ;Ｓｌｏｍｃｚｙｎ ｓｋｉ等[8] 研究了Ｂｏｔｒｙｔｉｓｃｉｎｅｒｅａ的漆酶特性 ;国内有关研究报道较少[3 ,4 ] ,筛选漆酶高…     

PHB颗粒在红豆草根瘤细菌发育中的动态变化

红豆草根瘤胞间隙和侵入线中另有个别细菌含有PHB颗粒,而且数量很少,一个细菌通常仅有一个。随着细菌被从侵入线中释放到寄主细胞中,这些PHB颗粒立即消失。幼龄细菌不含PHB颗粒,成熟细菌一般也不含这种内含物。当细菌衰老时,它们又再度出现,并大量增加,而后很快减少,直至完全消失。从未发现这种颗粒存在于解体细菌中,尽管它们处于各种不同的解体状态。PHB颗粒在细菌发育中的变化表明,它的多少不仅与根瘤细菌发育密切有关,而且也受制于根瘤品种。     

利用原生质体技术选育聚β—羟基丁酸高产菌株的研究初报

从城市生活污水中分离筛选到1株浮游球衣细菌（Sphaerotilus natans)。以浮游球细菌G-8菌为出发菌株,采用甘氨酸-溶菌酶-EDTA方法,研究了各种因素对原生质体形成和再生的影响。在原生质体形成和再生的最佳条件下制备原生质体,G-8菌原生质体形成率和再生率分别达96.8%和29.8%。该菌株能在自然pH(pH6-7)和29℃条件下良好生长,细胞可大量积累聚β-羟基丁酸（PHB）,从对数期到稳定期,该菌积累的PHB量可达68g/L。     

富养罗尔斯通氏菌菌株PHB"泄漏"机理的初探

建立了一种分离纯化聚羟基丁酸(Polyhydroxybutyrate,PHB)颗粒的改良方法。采用这种方法从Ralstonia eutropha菌株H16(野生型)、SK1489(Tn5诱变的PHB泄漏菌株)、JMP222(野生的PHB泄漏菌株)分离了PHB颗粒。进一步比较研究了不同菌株的PHB解聚酶和3-羟基丁酸脱氢酶的活性。研究结果表明,菌株SK1489的PHB解聚酶活性(48h培养后达1．82U／mg)明显高于野生型菌株H16(48h培养后达0．37U／mg),菌株JMP222的3-羟基丁酸脱氢酶活性(培养96h后达165．9U／mg)比菌株H16培养(96h后达64．0U／mg)高许多。这些结果显示,不同菌株PHB的泄漏有不同的原因,突变株SK1489导致PHB泄漏的原因是解聚酶活性高,而野生型JMP222PHB泄漏的原因主要是3-羟基丁酸脱氢酶活性高。     

用β-紫罗兰酮筛选虾青素高产菌株

用β-紫罗兰酮作为筛选剂选择性分离海洋红酵母虾青素高产突变菌株.实验结果表明,在β-紫罗兰酮存在的情况下,由于类胡萝卜素合成受到抑制,海洋红酵母的生物量和虾青素合成量都减少;当平板培养基中β-紫罗兰酮浓度达到370 mg/L时,海洋红酵母的致死率为92.3%:在这种平板培养基上涂布经甲基磺酸乙酯诱变的海洋红酵母,随机筛选200个菌落,结果表明生物量、虾青素体积产率和细胞产率均有所提高的正突变株占18%,生物量、虾青素体积产率和细胞产率的单项指标有所提高的正突变株分别占22.5%、45%和46%.该实验结果表明在分离培养基中添加β-紫罗兰酮可选择性地分离海洋红酵母虾青素高产菌株,提高虾青素高产突变株的筛选效率.     

Physical and chemical characteristics of the Blue Nile and the White Nile at Khartoum

Fortnightly measurements of physical and chemical variables were made at two locations on the Blue and White Niles near Khartoum from August 1968 to December 1970. Variables analysed from each river were: temperature, pH, total residue, current velocity, oxygen, alkalinity, phosphate, nitrate, ammonia, silica, sulphate, iron, calcium, magnesium, sodium, potassium and oxidizable organic matter. The seasonal variations of these factors in the two Niles are compared and the interrelationships existing between some of them are discussed. Comparisons with earlier studies on the Nile and with some tropical rivers are made.In the Blue Nile, the amounts of suspended matter and nutrients are largely dependent upon the flood regime. Nitrate, phosphate, iron, oxidizable organic matter and total residue increase considerably in the Blue Nile when the river is in flood (peaks: 1 880 µg NO₃-N l^–1; 0.31 mg Fe l^–1; 3 842 mg total residue · l^–1).In the White Nile, concentrations of nitrate, phosphate, iron, oxidizable organic matter and total residue attain their peaks during the rainy season (270 µg NO₃-N l^–1; 163 tag PO₄-P l^–1; 0.46 mg Fe · l^–1; 502 mg total residue · l^–1).In both rivers, alkalinity, calcium, sodium and potassium tend to increase during the dry season while declining in the rainy season. Silica is depleted at certain times of the year, yet relatively high concentrations are maintained throughout the year and were not expected to limit growth of diatoms. Fall in silica concentrations, unlike nitrate, phosphate and iron, was always followed by a rapid restoration of a high level. Silica and magnesium showed no response to changes in discharge rates.     

丹参冠瘿组织高产株系选择和丹参酮的产生

丹参(Salvia miltiorrhiza)是治疗心血管系统疾病的重要中草药。前文我们已报道了丹参冠瘿组织培养的部分研究结果^[1].利用根癌农杆菌(Agrobacterium tumefaciens)Ti 质粒转化产生的冠瘿组织进行高产株系选择的研究还很少,本文报道丹参冠瘿组织高产株系选择和丹参酮产生的研究结果。     

尼罗红荧光优化法快速检测微拟球藻细胞内油脂含量

【目的】利用尼罗红荧光染色法快速检测微拟球藻细胞内的油脂含量。【方法】系统性地调整激发波长与发射波长,确定最佳二甲基亚砜(DMSO)浓度、尼罗红终浓度、染色时间和细胞密度的范围,比较重量法和Triolein标准品的油脂含量分别与荧光强度之间的关系。【结果】分析得出了尼罗红荧光强度与微拟球藻细胞油脂含量的关系,确定优化后染色条件为:二甲基亚砜浓度为5%,尼罗红终浓度为1 mg/L,染色时间为6 min,且细胞密度为(0.5-3.0)×10~6 cells/m L的范围内,激发波长和发射波长分别为515 nm和570 nm。重量法和Triolein标准品的油脂含量分别与尼罗红染色荧光强度之间的关联性,表明尼罗红荧光染色方法可以用来快速准确地检测细胞内的油脂含量,且油脂含量与荧光强度之间正相关,相关系数R~2为0.997 3。尼罗红染色优化后油脂的检测下限达到2μg,大大减少了测定油脂含量所需细胞量。【结论】针对不同种属系统性地确认了荧光激发波长和发射波长,并优化验证得到了最佳尼罗红荧光染色条件,可以快速准确地检测微量微拟球藻细胞内的油脂含量,便于大规模筛选高产油突变藻株。     

用RAPD技术对罗非鱼遗传变异的研究及应用(英文)

应用随机扩增多态性ＤＮＡ（ＲＡＰＤ）技术检测了一个奥利亚罗非鱼（ａｕ）和湘湖（ｎｘ）、美国（ｎｍ）、沙市（ｎｐ）三个尼罗罗非鱼养殖群体（Ｔａｂｌｅ１）。在２０个引物（Ｔａｂｌｅ ２）中筛选到１２个引物,它们的扩增产物显示了罗非鱼和尼罗罗非鱼二者在群体内或群体间存在遗传差异。其中（Ｆｉｇ．１）, ＯＰＺ０６、 ＯＰＺ１６、 ＯＰＺ１２和 ＯＰＺ１９四个引物分别有一个扩增片段具有种的特异性。它们的大小分别是９００、１５００、１７００和７３０ｂｐ。可以作为鉴别罗非鱼和尼罗罗非鱼二者的分子遗传标记。湘湖（ｎｘ）、美国（ｎｍ）和沙市（ｎｐ）三个尼罗罗非鱼群体内遗传变异相似性指数Ｓ分别为 ０．７９８、 ０．７９５和０．８２４（Ｔａｂｌｅ ３）。表明：这三个尼罗罗非鱼群体都保留了较高水平的遗传变异。而奥利亚罗非鱼（ａｕ）的群体内遗传变异最小。奥利亚罗非鱼（ａｕ）与湘湖（ｎｘ）、美国（ｕｍ）、沙市（ｕｐ）三个尼罗罗非鱼群体之间的遗传距离分别是０．２８５、０．２６２和０．３４４（Ｔａｂｌｅ ３）,说明奥利亚罗非鱼（ａｕ）和沙市尼罗罗非鱼（ｎｐ）杂交将可能产生较强的杂种优势。     

Energetics of RNA binding by the West Nile virus RNA triphosphatase

The West Nile virus (WNV) RNA genome harbors the characteristic methylated cap structure present at the 5' end of eukaryotic mRNAs. In the present study, we report a detailed study of the binding energetics and thermodynamic parameters involved in the interaction between RNA and the WNV RNA triphosphatase, an enzyme involved in the synthesis of the RNA cap structure. Fluorescence spectroscopy assays revealed that the initial interaction between RNA and the enzyme is characterized by a high enthalpy of association and that the minimal RNA binding site of NS3 is 13 nucleotides. In order to provide insight into the relationship between the enzyme structure and RNA binding, we also correlated the effect of RNA binding on protein structure using both circular dichroism and denaturation studies as structural indicators. Our data indicate that the protein undergoes structural modifications upon RNA binding, although the interaction does not significantly modify the stability of the protein.     

我国新引进吉富品系尼罗罗非鱼群体的遗传多样性分析

选取罗非鱼(Oreochromis spp.)第二代遗传连锁图谱中的26个微卫星位点,对淡水渔业研究中心引进的、由60个家系组成的吉富品系尼罗罗非鱼(O.niloticus)群体进行遗传结构分析。结果显示,26个微卫星位点在吉富罗非鱼群体中共检测到124个等位基因,各位点的等位基因数为3～7个,平均4.8个。片段长度104～322 bp,平均杂合度观测值为0.622 1,平均杂合度期望值为0.642 3,平均多态信息含量(PIC)为0.633 4。所检测的26个位点中,有25个位点属于高度多态位点(PIC0.5),占所检测位点的96.15%;1个位点属于中度多态位点。结果表明,该吉富罗非鱼群体多态信息含量丰富,遗传多样性水平较高。因而该群体仍然具有较大的选育潜力,可以作为选育的基础群体开展进一步的选育工作。     

Effects of sedimentation on the storage capacity of the High Aswan Dam reservoir

The River Nile receives most of its sediment load from the Atbara and Blue Nile rivers, which carry eroded sediments north from the Ethiopian mountains during the seasonal flood period between August and October. Prior to the construction and operation of the High Aswan Dam, in 1964, 9–10 × 10⁶ metric tons of suspended sediment were deposited annually in the flood plain of the Nile, while about 93% of the total average annual suspended load of 124 × 10⁶ metric tons was carried out into the Mediterranean Sea. Since the full operation of the High Aswan Dam in 1968, the flood discharge of the Nile, below the dam, has been greatly modified and more than 98% of the total suspended load has been retained within the reservoir. Based on long-term records; estimated relationships between discharge and suspended load, and field measurements, the life span of the dead storage capacity has been estimated at a minimum of about 360 years. Although this preliminary calculation is less than the estimated design capacity of 450 years, it is expected that progressively more suspended solids will be released in the outflow of the reservoir and that together with the use of flood diversion schemes the High Aswan Dam is likely to approach its design life span.     

The food and growth of pelagic 0+ Alestes nurse (Characidae) from the Blue Nile,near Khartoum

The food, feeding habits and growth of 0+ Alestes nurse (Rüppel) from the Blue Nile near Khartoum, was investigated. 0+ A. nurse were observed to change their food from phytoplankton and small zooplankton during their pre-metamorphosis stages, to larger zooplankton (Daphnia spp., Chydorus sp.) and insects (larvae, pupae and imagines) during their post-metamorphosis stage.Their growth in length and weight was characterized by two distinct stanzas; during the first stanza, the fish increased in total length at a rate of 0.196 mm mm^–1 wk^–1, and in wet weight at a rate of 0.430 g g^–1 wk^–1. These rates were reduced during the second stanza to 0.046 mm mm^–1 and 0.080 g g^–1 wk^–1, respectively. The calculated value of the exponent b in their length-weight relationship suggested that their growth was allometric.     

遗传改良"新吉富" (NEW GIFT)尼罗罗非鱼RAPD-SCAR标记开发及其在品系鉴别中的应用

“新吉富”(NEW GIFT)尼罗罗非鱼是以1994年引进的 “吉富”(GIFT)品系尼罗罗非鱼为基础群体,经过14年9代系统选育后获得的新品种,该品种已被国家审定为良种,已在全国推广,迫切需要有效的分子遗传标记来鉴别该新品种并实施科学管理。该研究对良种群体开展了RAPD分析,并进一步将特异性的RAPD标记转化成了稳定的SCAR标记。 通过对“新吉富”群体的RAPD分析,找到了两个品种特异性的RAPD条带（S₃₀₄^{624 bp}和S₃₆^{568 bp}）。对品种特异性的RAPD条带进行了胶回收、克隆和序列测定。根据测序结果设计了两对位点特异性引物来扩增这两个品种特异性条带（简称SCAR标记Ⅰ和Ⅱ）,检测这两个SCAR标记在“新吉富”、“吉富”以及国内外7个养殖品系中的出现频率。SCAR标记Ⅰ（553 bp）在“新吉富”群体中的出现频率为85.7%,而在其基础群体（“吉富”）中的出现频率仅为16.7%;SCAR标记Ⅱ（558 bp）在“新吉富”群体中的出现频率达到91.4%,但在其它7个养殖品系中的出现频率仅在0%～70%之间。为验证这两个标记的可靠性,检测了这两个标记在一个埃及罗非鱼野生群体中的出现频率,发现SCAR标记Ⅰ和SCAR标记Ⅱ的出现频率分别为10%和70%,远低于在“新吉富”群体中的出现频率。这两个SCAR标记在“新吉富”良种群体中的高出现频率,预示它们可能与生长性能相关QTL位点间存在连锁关系,显示SCAR标记技术对选育世代间的纵向追溯、以及选育良种群体与其它养殖品系间的横向鉴别有较好应用前景。     

Study of the Effect of Different Techniques on Diversity of Freshwater Hyphomycetes in the River Nile (Upper Egypt)

Four different techniques were applied, lead mapping of Eucalptus rostrata; randomly leaf sampling; Millipore filtration; spores in foam, for the study of aquatic hyphomycets communities in the River Nile. Triscelophorus monlosporus, Anguilospora longissima, Flagellospora curvula and Tetracladium marchalianum were the dominant species in all the techniques used. Aquatic hyphomycetes diversity was high by using leaf mapping (23 species) and randomly leaf sampling (30 species) but it was low by using Millipore filtration (11 species) and spores in foam (8 species) techniques.