联合固氮菌Enteroabcter gergoviae 57-7氮代谢相关基因的克隆和泌氨工程菌的构建

从构建具有稳定泌氨能力的联合固氮工程菌的目的出发，首先构建Enterobacter gergoviae57-7 (E57-7)基因文库并与所筛选的泌氨突变株进行遗传互补实验，得到可互补 泌氨特性的克隆，经Southern杂交后推测其中包含与glnA、amtB基因无关的另一类与泌 氨相关的基因。同时根据铵载体基因amtB的已知序列设计两对简并性引物，经PCR从 E57-7 DNA中扩增得到约340bp的片段，序列分析和Blast序列同源性比较后确定为amtB 基因片段，申报并获得序列号AJ132232，最终从基因库中筛选到两个包含E57-7 amtB 基因的克隆。 用K. pneumoniae的glnA基因片段为探针，通过Southern杂交从E57-7基因库中筛选到包含有glnAntrBC基因的克隆，经亚克隆后对包含有这个操纵元的4316bp片段进行了全序列分析，申报GenBank获得序列号AF072440。在体外实验中构建了Km-cassette 插入glnA的重组质粒pA，将此质粒转入E57-7野生型菌株后经筛选同源重组子获得glnA 突变的具有稳定泌氨能力的菌株15、I9。并进行了盆栽玉米接种实验，确定在灭菌上壤实验体系下I5对玉米幼苗有显著促生效应。 利用绿色荧光蛋白(GFP-S65T，V68L，S72A)基因建立分子生物学研究手段，构建了新 型克隆载体pGreenLD，建立了绿白斑筛选重组质粒的的技术。构建组成型表达gfp的质粒载体研究了E57-7在玉米根际的定殖模式;构建nifH-gfp表达载体，确定在与植物联合生活时其固氮酶结构基因nifHDK的表达与碳源物质供应密切相关。利用不同抗性基因和gfp基因片段构建出在E57-7中组成型表达抗性和GFP的质粒载体，建立了监测接种菌在土壤中释放的双标记系统。 最后克隆了E57-7 glg cluster并测定部分glgCA和glgP基因序列，申报后获得序列号AJ132233和AJ132234，这是首例从联合固氮菌中克隆得到glg cluster的报道。

几种固氮蓝藻的固氮酶活性及其某些特性

对三种固氮蓝藻:固氮鱼腥藻(水生686)、柱孢鱼腥藻和鱼腥藻7120的整细胞及无细胞抽提液的固氮酶活性,进行了比较研究。水生686的整细胞酶活虽然不低(51.9毫米乙烯峰高/光密度/30分),仅次于柱孢鱼腥藻,但其无细胞抽提液的酶活却最低。这可能与它含有大量藻胶有关。研究了Mn~(++)、Fe~(++)对蓝藻固氮酶的作用,以及测定其在不同酶浓度下的反应动力学表明:柱孢鱼腥藻中不存在象深红螺菌中所看到的那种激活因子。用甲苯-乙醇溶液处理藻细胞,对固氮酶作原位测定,探索了它的氧损伤及氧保护机理。

Photoreduction of mercuric salt in UV/Fe (III)-oxalate complex system

In this paper, the photochemical reduction process of Hg (II) in aqueous solution containing ferric iron and oxalate (Ox) has been studied. Under the radiation of a low-pressure mercury lamp (lambda = 253.7 nm, 8W), Fe(III)-oxalate complexes undergo photolysis to produce ferrous ions and other organic reductive species, which reduce Hg(II) subsequently. For 0.1 mg/L Hg (II), the photoreduction efficiency is comparatively higher in the solution at pH 5.0 than that over the range of 3.0 similar to 8.0. The photoreduction efficiency of Ho (II) in aqueous solution increases with increasing, initial concentration of ferric ions from 0.02 mmol/L to 0.2 mmol/L and initial concentration of oxalate from 0.96 mmol/L to 4.8 mmol/L and then gradually approaches to a steady state. CH3OH also contributes the reduction of Hg (II). We investigate the increase of the ferric, oxalate and CH3OH concentrations resulting from the increase of reduction efficiency of Hg (II). It can be seen that ferrous ions and other reactive species are reductants of Hg (II), and the reaction product with oxalate is mainly volatile metallic mercury.

Study on fluorescence emission and synchronous-scan fluorescence spectrea of Nitzschia hantzschiana solution with FE(III)

The characterization of the algal Nitzschia hantzschiana solution with (or without) Fe(III) was carried out using fluorescence emission and synchronous-scan spectroscopy. An emission peak (excited at 440 nm) was observed at 675 nm for Nitzschia hantzschiana solution. The effective characterization method used was synchronous-scan fluorescence spectroscopy (SFS). A wavelength difference (Delta lambda) of 90 nm was maintained between excitation and emission wavelengths. The peak was observed at about 236(ex) nm (326(em) nm) for synchronous fluorescence spectroscopy. Fe(III) was an effective quencher. The relationship between I-0/I (quenching efficiency) and c (concentration of Fe (III) added) was a linear correlation for the algal solution with Fe(III). Effects of pH on synchronous-scan fluorescence intensity were evident.

Preliminary study on the photoproduction of hydroxyl radicals in aqueous solution with Aldrich humic acid, algae and Fe(III) under high-pressure mercury lamp irradiation

Under a high-pressure mercury lamp (HPML) and using an exposure time of 4 h, the photoproduction of hydroxyl radicals ((OH)-O-.) could be induced in an aqueous solution containing humic acid (HA). Hydroxyl radicals were determined by high-performance liquid chromatography using benzene as a probe. The results showed that (OH)-O-. photoproduction increased from 1.80 to 2.74 muM by increasing the HA concentration from 10 to 40 mg L-1 at an exposure time of 4 h (pH 6.5). Hydroxyl radical photoproduction in aqueous solutions of HA containing algae was greater than that in the aqueous solutions of HA without algae. The photoproduction of (OH)-O-. in the HA solution with Fe(111) was greater than that of the solution without Fe(III) at pH ranging from 4.0 to 8.0. The photoproduction of (OH)-O-. in HA solution with algae with or without Fe(111) under a 250 W HPML was greater than that under a 125 W HPML. The photoproduction of (OH)-O-. in irradiated samples was influenced by the pH. The results showed that HPML exposure for 4 h in the 4-8 pH range led to the highest (OH)-O-. photoproduction at pH 4.0.

Energy budget of Nile tilapia (Oreochromis niloticus) in relation to ration size

Nile tilapia weighing 8.29-11.02 g were fed a practical diet at seven ration levels (starvation, 0.5, 1, 2, 3, 4% body weight per day and satiation) twice a day at 30 degrees C. Feed consumption, apparent digestibility, nitrogenous excretion and growth were determined directly, and heat production was calculated by difference of energy budget. The relationship between specific growth rate in wet weight (SGR(w), percentage per day) and ration size (RL, percentage per day) was a decelerating curve described as SGR(w) = 2.98 (1 - e(-0.61(RL-0.43))). The apparent digestibility coefficients for dry matter and protein showed a decreasing pattern with increasing ration while the apparent digestibility coefficient of energy was not significantly affected by ration size. The proportion of gross energy intake lost in nitrogenous excretion tended to decrease with increasing ration. Feed efficiency was highest, and the proportion of gross energy intake channelled to heat production was lowest, at an intermediate ration level (2% per day). The energy budget at the satiation level was: 100IE = 16.9FE + 1.2(ZE + UE) + 62.3HE + 19.6RE, where IE, FE, (ZE + UE), HE and RE represent gross energy intake, faecal energy, excretory (non-faecal) energy loss, heat production and recovered energy (growth), respectively. (C) 1997 Elsevier Science B.V.

EFFECT OF RATION SIZE ON THE GROWTH AND ENERGY BUDGET OF THE GRASS CARP, CTENOPHARYNGODON-IDELLA VAL

Young grass carp (12-13 g) were kept at five ration levels ranging from starvation to ad libitum feeding at 30-degrees-C. They were fed duckweed. Food consumption, absorption efficiency and growth were determined directly, and metabolism and nitrogenous excretion calculated indirectly from energy and nitrogen budgets, respectively. The relationship between specific growth rate and ration size was linear. Absorption efficiency for energy was not affected by ration size and averaged 50.6 +/- 0.57% (mean +/- s.e.). Depending on ration size, energy lost in excretion accounted for 4.5-5.9% of the food energy, energy channelled to metabolism accounted for 34.4-48.3% of the food energy, and energy retained as growth accounted for 6.7-11.9% of the food energy. Regardless of ration, a constant proportion of food energy (30.7%) was accounted for by feeding metabolism (total metabolism minus fasting metabolism). The energy budget at the maximum ration was: 100 C = 49.1F + 4.5U + 3.6R(fa) + 30.9R(fe) + 11.9G, where C, F, U, R(fa), R(fe) and G represent food consumption, faecal production, excretion, fasting metabolism, feeding metabolism and growth, respectively.

InP中的H、Fe和Cu的研究

Submitted by zhangdi (zhangdi@red.semi.ac.cn) on 2009-04-13T11:45:31Z

Fe/Al2O3/GaAs 异质结构中自旋相关输运性质的研究

Submitted by zhangdi (zhangdi@red.semi.ac.cn) on 2009-04-13T11:45:31Z