一种重组人α1型干扰素突变体IFN-α1/86D的纯化与鉴定

本文研究了重组人α1型干扰素突变体IFN-α1/86D在摇瓶培养和分批发酵培养的重组大肠杆菌DH5α株中的表达动态,并采用阳离子交换层析和抗α1型干扰素单克隆抗体亲和层析的两步流程对其进行了纯化,得到SDS-PAGE纯及高效液相层析(HPLC)纯的IFN-α1/86D产品,共比活性为2.3×10~7单位/毫克蛋白。N端氨基酸序列测定的结果表明,纯化产品的纯度在95％以上,但其N端不均一,产品中含有两种N端序列正确的IFN-α1/86D活性分子,即约75％为去Met分子和约25％为带Met分子。     

鱼腥藻1017株的混合营养型生长

鱼腥藻１０１７株混合营养型生长有其特点,外源葡萄糖对生长的刺激不仅在低光强（８００ｌｘ）,而且在高光强（７０００ｌｘ）也表现出来。其混合营养型生长速率在８００－７０００ｌｘ范围内随光照强度的增加而增加,在葡萄糖浓度５－２０ｍｍｏｌ／Ｌ范围内随外源葡萄糖浓度增加而增加。鱼腥藻１０１７株混合营养型生长与光能自养生长相比,生长速率明显提高,对数生长期延长,收获物浓度显著增高,生物量显著提高。     

鸡减蛋综合征病毒（EDSV—76）末端前体蛋白的基因结构分析

从中国发病鸡群中分离的鸡减蛋综合征病毒弱毒株ＡＡ－２,经常规方法提取其病毒核酸后,组建了完整的限制性内切酶ＰｓｔＩ及ＨｉｎｇⅢ水解片段的基因文库,并对其中ＨｉｎｄⅢ,－ＳａｃⅠ进行了序列测定。同源比较分析证明：其Ｌ链含编码病毒末端前体蛋白,容量为５８０个氨基酸残基的开放读码框架。     

人参茎叶皂甙对高胆固醇饮食大鼠再灌注性心律失常和脂质过氧化的影响(英文)

研究人参茎叶皂甙(GSL)对高胆固醇饮食大鼠心肌再灌注性心律失常(RPA_r)和脂质过氧化的影响。方法:将胆固醇乳剂用灌胃法饲养大鼠14d,建立高脂血症模型,各组大鼠进行心肌缺血再灌注实验,观察高脂血症和GSL对大鼠心肌缺血再灌注2h后血丙二醇(MDA),超氧化物歧化酶(SOD)和一氧化氮(NO)水平的影响和对再灌注性心律失常发生率的影响。结果显示:(1)用胆固醇乳剂饲养大鼠14d,成功建立高脂血症模型。同时给予GSL14d有明显降脂作用。(2)高脂血症状态下,心肌缺血再灌注2h后,血MDA升高(p<0.01),SOD降低(p<0.01)和NO(p<0.05)降低,再灌注10min内RPAr的发生率增高。(3)GSL组再灌注后2h的血MDA降低,而SOD和NO水平显著升高;使RPAr发生率大为降低,无VF发生。实验显示高脂血症加重心肌缺血再灌注损伤和提高RPAr发生率及动物死亡率,GSL可减少高脂饮食大鼠脂质过氧化和诱导体内NO生成而减轻缺血再灌注心肌损伤,降低缺血再灌注性心律失常发生率。     

不同发酵条件下产甘油假丝酵母有机酸代谢的研究

产甘油假丝酵母 (Candidaglycerolgenesis)发酵产生的有机酸对丙三醇产品质量和产率均有影响。发现在发酵其它条件恒定 ,装液比和玉米浆浓度增加时 ,发酵液总酸是递增的。在装液比为 0 2和玉米浆浓度为 8g L时 ,丙酮酸和乳酸在细胞生长期可分别积累达 4 1g L和 1 0g L ,比正常发酵时增加 2倍以上 ,丙三醇产率也低 ;然而 ,装液比为 0 0 8和玉米浆浓度为 4g L时 ,丙酮酸和乳酸产生较低 ,丙三醇产率较高 ,但乙酸积累比供氧不足时高 ,可达 2 6g L。发酵过程中有机酸被细胞代谢 ,含量逐渐下降 ,如在初糖浓度为 1 0 0g L时 ,有机酸在细胞生长期积累至高峰后 ,丙三醇和有机酸随之均降低至较低含量 ,并且丙酮酸或乳酸可以转化为乙酸。此外 ,在外加一些添加剂时对其产生有机酸也有影响 ,如添加 1 %油酸和VB1时可以降低乙酸的积累 ,同时增加丙酮酸的含量 ,丙三醇产量也有所增加 ;而丙酮酸结构类似物氟代丙酮酸和亚硫酸盐促进乙酸的产生 ,使酮戊二酸合成减少 ,丙三醇产量约增加 2 0 %。     

黄瓜转新型抗菌蛋白基因GNK2-1及其抗枯萎病的研究

GNK2-1为一种来自银杏（Ginkgo biloba）种仁的新型抗真菌蛋白,具有较强的真菌抗性且性质稳定。序列分析表明,其结构与所有已知的抗真菌蛋白不同,而与富含半胱氨酸的植物类受体激酶的胞外结构域相似。为探索GNK2-1基因在黄瓜（Cucumis sativus）抗病反应中的作用,利用基因重组技术构建了GNK2-1的高效组成型表达载体,并利用根癌农杆菌（Agrobacterium tumefaciens）介导转入黄瓜栽培品种农城3号（Cucumis sativus＇ Nongcheng No.3＇）基因组中。通过对获得的抗性植株进行PCR、RT-PCR和Western blot检测分析,结果表明GNK2-1基因可在T0代转基因植株中转录表达,并能在T1代转基因黄瓜中稳定遗传。离体枯萎病抗性鉴定结果表明,转GNK2-1基因的黄瓜对枯萎病的抗性增强,GNK2-1可以作为黄瓜抗病性改良的潜在基因资源。     

Bacteria abundance and diversity of different life stages of Plutella xylostella (Lepidoptera: Plutellidae), revealed by bacteria culture‐dependent and PCR‐DGGE methods

Microbial abundance and diversity of different life stages (fourth instar larvae, pupae and adults) of the diamondback moth, Plutella xylostella L., collected from field and reared in laboratory, were investigated using bacteria culture‐dependent method and PCR‐DGGE analysis based on the sequence of bacteria 16S rRNA V3 region gene. A large quantity of bacteria was found in all life stages of P. xylostella. Field population had higher quantity of bacteria than laboratory population, and larval gut had higher quantity than pupae and adults. Culturable bacteria differed in different life stages of P. xylostella. Twenty‐five different bacterial strains were identified in total, among them 20 strains were presented in larval gut, only 8 strains in pupae and 14 strains in adults were detected. Firmicutes bacteria, Bacillus sp., were the most dominant species in every life stage. 15 distinct bands were obtained from DGGE electrophoresis gel. The sequences blasted in GenBank database showed these bacteria belonged to six different genera. Phylogenetic analysis showed the sequences of the bacteria belonged to the Actinobacteri, Proteobacteria and Firmicutes. Serratia sp. in Proteobacteria was the most abundant species in larval gut. In pupae, unculturable bacteria were the most dominant species, and unculturable bacteria and Serratia sp. were the most dominant species in adults. Our study suggested that a combination of molecular and traditional culturing methods can be effectively used to analyze and to determine the diversity of gut microflora. These known bacteria may play important roles in development of P. xylostella.     

乳酸菌黏附小鼠派伊尔结及影响因子的研究

目的研究乳酸菌结合小鼠派伊尔结（PP）的性质,确定乳酸菌黏附小鼠PP的影响因子。方法测定10株荧光标记的乳酸菌对PP组织切片黏附性,分析细菌表面疏水性和酵母、红细胞凝集性与黏附性的关系。结果凝集酵母、红细胞能力强的乳酸菌,PP黏附性强。除嗜酸乳杆菌（L.acidophilm）外,其他9株乳酸菌的表面疏水性与凝集酵母、红细胞的能力呈显著的负相关性（r=-0．60和r=-0．53,P〈0．05）。L.acidophilm黏附PP的能力最强。L．acidophilm黏附PP能力和红细胞凝集性均可被D（＋）-甘露糖、D（＋）-半乳糖抑制。结论乳酸菌黏附PP的能力受疏水性和表面特异性凝集素的影响。L.acidophilm表面D（＋）-甘露糖、D（＋）-半乳糖特异性凝集素可能参与黏附PP的过程。     

Delayed development of the whitefly (Bemisia tabaci) and increased parasitism by Encarsia bimaculata in response to sublethal doses of piperonyl butoxide

Abstract Effects of sublethal piperonyl butoxide (PB) on parasitization of Bemisia tabaci (Gennadius ) (Hemiptera: Aleyrodidae) by Encarsia bimaculata Heraty et Polaszek (Hymenoptera: Aphelinidae) were evaluated both in cage and greenhouse experiments. When first, second and third instar B. tabaci nymphs were treated with PB, all but the first instar were significantly prolonged. Data indicated that sublethal PB could improve E. bimaculata parasitism rates without influencing parasitoid eclosion rates. Prolonged development increased rates of parasitism by E. bimaculata, from 17.6% to 24.7% in cages, presumably by increasing the duration of host exposure. Sublethal PB combined with E. bimaculata as an integrated approach to control B. tabaci was evaluated using life table parameters under greenhouse conditions. Indices of population trend (I) calculated from life tables were estimated at 4.6 for B. tabaci exposed to PB and parasitoids compared to 14.1 with parasitoids alone and 23.5 in untreated controls. The results showed that after PB was sprayed and parasitoids introduced, development of B. tabaci was delayed and the peak of each stage was postponed. The older nymphal stage had highest mortality, primarily due to mortality caused by parasitism by E. bimaculata.     

Reduction in glutamate uptake is associated with extrasynaptic NMDA and metabotropic glutamate receptor activation at the hippocampal CA1 synapse of aged rats

This study aims to determine whether the regulation of extracellular glutamate is altered during aging and its possible consequences on synaptic transmission and plasticity. A decrease in the expression of the glial glutamate transporters GLAST and GLT‐1 and reduced glutamate uptake occur in the aged (24–27 months) Sprague–Dawley rat hippocampus. Glutamatergic excitatory postsynaptic potentials recorded extracellularly in ex vivo hippocampal slices from adult (3–5 months) and aged rats are depressed by DL‐TBOA, an inhibitor of glutamate transporter activity, in an N‐Methyl‐d‐ Aspartate (NMDA)‐receptor‐dependent manner. In aged but not in young rats, part of the depressing effect of DL‐TBOA also involves metabotropic glutamate receptor (mGluRs) activation as it is significantly reduced by the specific mGluR antagonist d‐methyl‐4‐carboxy‐phenylglycine (MCPG). The paired‐pulse facilitation ratio, a functional index of glutamate release, is reduced by MCPG in aged slices to a level comparable to that in young rats both under control conditions and after being enhanced by DL‐TBOA. These results suggest that the age‐associated glutamate uptake deficiency favors presynaptic mGluR activation that lowers glutamate release. In parallel, 2 Hz‐induced long‐term depression is significantly decreased in aged animals and is fully restored by MCPG. All these data indicate a facilitated activation of extrasynaptic NMDAR and mGluRs in aged rats, possibly because of an altered distribution of glutamate in the extrasynaptic space. This in turn affects synaptic transmission and plasticity within the aged hippocampal CA1 network.