Hydrogen production by combining two types of photosynthetic bacteria with different characteristics

The double-layer photobioreactor using two types of photosynthetic bacteria, Rhodobacter sphaeroides RV and its reduced-pigment mutant, MTP4, was developed for efficient hydrogen production. The two types of bacteria had different characteristics on light energy, hydrogen production rate and conversion efficiency. MTP4 produced hydrogen more efficiently under high light conditions and RV did so under low light conditions. Illuminated light toward the surface of a photobioreactor quasi-exponentially declines as it penetrates into the reactor. When two types of bacteria were placed using the developed reactor according to this light distribution, the hydrogen production rate reached 3.64 l/m²/h at a light intensity of 500 W/m² in 24 h and the conversion efficiency of light energy to hydrogen was 2.18%. These values were 33% higher than those of only using RV. The low light in the deep part of the reactor was utilized efficiently, resulting in a higher hydrogen production rate.     

乙型肝炎病毒s基因变异株抗原表位的研究

对HepG2细胞表达的HBsAg的点突变体129Ha、142Lf、145Rb、143Sg和148及野生型HBsAgb3进行了初步纯化及抗原表位的研究，发现这些突变体的颗粒密度与野生型HBsAg差异无显著意义，约为1．2；用5种HBsAga决定簇的单克隆抗体对其表位进行分析，发现129Ha、142Lf、145Rb和142Sg与这5种单抗的结合力与野生型HBsAg差异大显著意义，但148突变体的表位与野生型HBsAg明显不同，其原因还有待进一步研究。     

睫状神经营养因子突变体的克隆、表达、纯化及生物学活性

目的设计具有更高生物学活性的睫状神经营养因子突变体,并对其进行表达、纯化及生物学活性检测。方法以计算机分子模拟系统设计突变体,重叠延伸PCR方法获得突变体的编码区DNA序列,克隆入表达载体pThioHisA,转化E.coliBL21,以IPTG诱导表达。复性纯化后,用鸡胚背根神经节无血清培养法和小鼠减重法进行生物学活性测定。结果目的蛋白以包涵体形式存在,表达量在35%以上,纯化后的纯度达95%以上,能有效地促进鸡胚背根神经节的生长,并能使正常小鼠的体重降低,脂肪指数下降。结论所设计的突变体经表达及纯化后,具有良好的生物学活性,为进一步研究突变体蛋白的促神经生长和减肥作用奠定了基础。     

刺桐胰蛋白酶抑制剂突变体在大肠杆菌中表达及在tPA纯化中的应用

目的构建刺桐胰蛋白酶抑制剂突变体(rserETI)原核表达载体,制备表达产物,用于tPA的纯化。方法设计合成rserETI编码序列DNA片段,以PCR法扩增出全长编码区序列,经酶切后克隆至pET9a表达载体,转化大肠杆菌JM109DE3,以IPTG诱导表达。表达产物经变性、复性、QFF层析纯化后,测定其活性,并与溴化氰活化的Sepharose偶联合成亲和层析柱,纯化rtPA。结果rserETI的表达量约占大肠杆菌菌体总蛋白的30%,复性率为80%,经QFF层析纯化后,蛋白浓度为1.58mg/ml,SDS-PAGE检测显示无杂蛋白污染,纯度大于95%,对rtPA突变体的抑制比活性为4×104IU/mg。偶联合成的rserETI-Sepharose亲和层析柱能特异性地纯化rtPA,rtPA突变体纯度达96%,比活性为5.07×105IU/mg。结论已成功构建了rserETI原核表达载体,并在大肠杆菌中高效表达,制备的表达产物可用于rtPA的纯化。     

Retention of inorganic arsenic by coryneform mutant strains

The natural resistance mechanisms of corynebacteria to respond to the environments containing high levels of arsenic were successfully adopted to develop inexpensive and selective extractants for submicrogram amounts of arsenic. Kinetic and equilibrium characteristics were evaluated, and a preliminary exploration of the capability of these strains to be used for arsenic speciation was also made in this work. Three kinetics models were used to fit the experimental data. It was found that the pseudo-first-order kinetics model was not quite adequate to describe the retention process, while the intraparticle diffusion and the pseudo-second-order kinetics models provide the best fits. The equilibrium isotherm showed that the retention of arsenic was consistent with the Langmuir equation and that the Freundlich and Dubinin-Radushkevich models provided poorer fits to the experimental data. The maximum effective retention capacity for arsenic was about 15.4 ng As/mg biomass. The amount of arsenic retained was directly measured in the biomass by forward planning a slurry electrothermal atomic absorption spectrometric procedure.     

假单胞菌phen8的降解特性及其缺失突变株的筛选

对分离筛选到的一株高效苯酚降解菌phen8（假单胞菌Pseudomonas sp.）的底物降解特性进行了分析,并筛选出四株缺的苯酚降解功能的突变株。结果表明,在苯酚初浓度为0－7mmol/L时,苯酚降解菌phen8的菌体密度和降率与培养基中的苯酚初浓度成正比,而大于7mmol/L的苯酚则明显地抑制降解反应,葡萄糖对苯酚的降解也表现出明显的抑制作用。与对照相比,在含有葡萄糖的基本培养基中培养20h后,菌体对苯酚的降解率减小47．2％。已筛选到的四个丢失苯酚降解功能的突变株,并今后降解基因的克隆和基因表达调控研究提供了良好的工作基础。     

利用细胞工程筛选烟草抗赤星病突变体的研究——一种双层培养基筛选系统的建立

为有效筛选烟草抗赤星病细胞突变体,本研究建立了一种双层培养基筛选系统。即在下层培养基中接种烟草赤星病(Alternaria alternata),在上层培养基中接种烤烟品种 NC89和净叶黄的花药(NC89高感赤星病,净叶黄高抗赤星病)。在26±1℃培养40天后发现,NC89的花药出苗率和花药平均出苗数与对照(不接种赤星病的培养基)相比,分别由30%和53颗降到2%和12颗,而净叶黄的花药出苗率和平均出苗数基本没有变化。对筛选出来的 NC89花培突变体进行了田间赤星病抗性鉴定,其中80%对赤星病抗性较 NC89有所提高,甚至部分接近高抗。这说明该双层培养基系统可以有效地对抗烟草赤星病的细胞突变体进行筛选。本系统尤其适应于(1)需长期培养;(2)毒素不易制备或易失活的病原菌的抗性筛选。     

Comparative Study of Fatty Acid Composition,Total Phenolics,and Antioxidant Capacity in Rapeseed Mutant Lines

There is limited variability within rapeseed germplasm in Morocco. Induced mutation was recently used to generate novel genetic variability and develop mutant lines combining desirable traits. In this context, nine promising advanced rapeseed M₂ mutant lines and the wild-type variety “INRA-CZH2” were evaluated for their seed oil content, fatty acid composition, total phenolic content (TPC), and free-radical scavenging activity (FRSA) by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) methods. The results showed significant variability among all mutants in seed oil content (38.14–42.04%) and fatty acids (SAFA = 5.49–10.99%, MUFA = 50.33–71.62%, PUFA = 22.89–8.68%). The mutant H2M-5 exhibited the highest fraction of MUFA and the lowest proportion of SAFA and PUFA, while the mutant H2M-4 showed the highest SAFA and PUFA amounts and the lowest MUFA level. TPC varied from 2.16 to 4.35 mg GAE/100 g. The highest amount was found in the mutant H2M-1, which is about twice that of other mutants and the wild-type variety. FRSA differed significantly among the samples, and the variations observed for DPPH and ABTS methods were 40.5–59.28% and 40.31–59.86%, respectively. FRSA was positively correlated to TPC in the sampled oils (r = 0.801 and 0.802, P < 0.01). This is the first report emphasizing the biochemical potential of rapeseed varieties and novel mutants in Morocco. H2M-1, H2M-4, and H2M-5 were proposed for the Rapeseed National Breeding Program, as they showed higher levels in some biochemical traits of interest.     

人蛋白C cDNA突变体的设计、克隆与序列分析

目的 通过对人蛋白C cDNA的定点突变,构建人蛋白C突变体cDNA,以期实现从哺乳动物细胞中直接表达出具有生物活性的人蛋白C产物。方法 针对人蛋白C cDNA序列设计引物以引入突变序列,采用逆转录聚合酶链反应(RT-PCR)和重组PCR方法,从人胎肝总RNA中分别钓取人蛋白C的重链和轻链,经重组PCR反应将两者连接,然后将其克隆入pGEM-T载体中,经酶切和PCR鉴定后进行测序。结果 经RT-PCR和重组PCR扩增获得大小为1374 hp的cDNA片段,并成功构建了人蛋白C cDNA突变体的克隆质粒pGEM-T/mhPC,序列分析证实所引入的编码8个氨基酸短肽的核苷酸序列突变位点正确取代了人蛋白C的活性肽,获得人蛋白C突变体cD-NA的克隆。结论 已成功进行了人蛋白C cDNA的突变,获得了人蛋白c cDNA突变体的克隆,为进一步进行人蛋白C突变体cDNA的表达和活性鉴定奠定了基础。     

Analysis of a Partial Male-Sterile Mutant of Arabidopsis thaliana Isolated from a Low-Energy Argon Ion Beam Mutagenized Pool

A screen for Arabidopsis fertility mutants, mutagenized by low-energy argon ion beam, yielded two partial male-sterile mutants tc243-1 and tc243-2 which have similar phenotypes. tc243-2 was investigated in detail. The segregation ratio of the mutant phenotypes in the M2 pools suggested that mutation behaved as single Mendelian recessive mutations, tc243 showed a series of mutant phenotypes, among which partial male-sterile was its striking mutant characteristic. Phenotype analysis indicates that there are four factors leading to male sterility, a. Floral organs normally develop inside the closed bud, but the anther filaments do not elongate sufficiently to position the locules above the stigma at anthesis, b. The anther locules do not dehisce at the time of flower opening （although limited dehiscence occurs later）, c. Pollens of mutant plants develop into several types of pollens at the trinucleated stage. as determined by staining with DAPI （4＇,6-diamidino-2-phenylindole）. which shows a variable size. shape and number of nucleus. d. The viability of pollens is lower than that of the wild type on the germination test in vivo and vitro.