固定化微生物降解土壤中菲和芘的研究

1　引　　言固定化微生物技术是 2 0世纪 70年代兴起的一种新型生物技术 ,目前已成为国内外学者的研究热点[8] ,现被广泛应用于处理化学工业废水 .其优点是可以大幅度提高参加反应微生物浓度 ;减少活性污泥数量 ;微生物被高分子材料包埋 ,耐环境冲击 ;根据需要选择有效微生物 ,可降低二次污染等特点 ,因而受到越来越多的关注[4,6 ,10 ] .包埋法是固定化技术最为普遍的使用方法 ,包埋载体的选择是固定化微生物的关键 ,理想的载体具有对微生物无毒性 ,传质性能好 ,性质稳定 ,不易被微生物分解 ,强度高寿命长和价格低廉等优点[2 ] .迄今 ,国内…     

胶体金免疫层析快速诊断技术

免疫胶体金技术是一种新的免疫学方法,该技术是一种将胶体金颗粒与包括抗原、抗体在内的许多蛋白质标记形成免疫金复合物的技术.目前在生物医学各领域得到了日益广泛的应用.在过去的十年里,基于膜上的快速诊断技术的发展为金标免疫试剂创造了巨大的市场.使用免疫金标技术制备的标记物作为检测系统中的指示试剂,应用在检测传染病、环境污染物、毒品、早孕以及兽医等领域.这两种技术的应用很大程度上提高了体外诊断技术的简便性和快速性.本文对其基本原理及其中应注意的问题做了一些介绍,并对其应用领域及发展方向进行了探讨.     

三七皂苷R1对大鼠缺血心肌VEGF、bFGF的影响

目的:探讨三七皂苷R1对大鼠缺血心肌VEGF、bFGF的影响。方法:选择雄性Wistar大鼠39只,建立心肌梗死(AMI)模型,术后24h存活大鼠随机分为药物组(n=13)、对照组(n=13),另设假手术组(n=8)。药物组给予三七皂苷R1水溶液(2.5 mg·kg-1·d-1)腹腔注射、对照组及假手术组给予等体积生理盐水腹腔注射,用药4周。于实验终点处死大鼠,心肌组织取材,Ⅷ因子染色计数微血管数(MVC)及微血管密度(MVD),免疫组织化学法观察缺血心肌VEGF、bFGF蛋白的表达。结果:药物组及对照组MVC、MVD均高于假手术组,且药物组高于对照组(P0.05);大鼠缺血心肌药物组及对照组VEGF、bFGF蛋白表达均高于假手术组(P0.05),且药物组高于对照组(P0.05)。结论:三七皂苷R1促进大鼠缺血心肌血管再生同时可上调缺血心肌VEGF、bFGF蛋白水平。     

牦牛卵泡细胞及其卵母细胞不同发育时期的结构变化

采集成年母牦牛卵巢,通过光镜和电镜对牦牛卵泡及其卵母细胞不同发育时期的结构变化进行了观察。结果发现当卵母细胞被单层立方卵泡细胞包围时,微绒毛开始出现,而皮质颗粒、透明带则在包被2-4层卵泡细胞时开始出现。随着卵母细胞的继续发育,透明带增厚,微绒毛由粗短变为细长,密度增加;皮质颗粒、线粒体、滑面内质网等细胞器的数目不断增加,并逐渐移行到质膜下;在移行的过程中,皮质颗粒成团存在。在囊状卵泡中,卵母细胞皮质颗粒呈线形分布于质膜下,线粒体、滑面内质网又移向胞质中央。卵母细胞借助微绒毛穿过透明带与卵泡细胞胞质突起相联系。结果证明牦牛卵泡和卵母细胞不同发育时期的结构变化与其它哺乳动物的基本相似。     

Phylogenetic distribution in the genus Mus of t-complex-specific DNA and protein markers: inferences on the origin of t-haplotypes

We have examined the phylogenetic distribution of two t-specific markers among representatives of various taxa belonging to the genus Mus. The centromeric TCP-1a marker (a testicular protein variant specific for all t-haplotypes so far studied) has also been apparently detected in several non-t representatives of the Mus IVA, Mus IVB, and probably M. cervicolor species. By contrast, a t-specific restriction- fragment-length polymorphism allele (RFLP) of the telomeric alpha- globin pseudogene DNA marker alpha-psi-4 was found only in animals belonging to the M. musculus-complex species either bearing genuine t- haplotypes or, like the M. m. bactrianus specimen studied here, likely to do so. This t-specific alpha-psi-4 RFLP allele was found to be as divergent from the RFLP alleles of the latter, non-t, taxonomical groups as it is from Mus 4A, Mus 4B, or M. spretus ones. These results suggest the presence of t-haplotypes and of t-specific markers in populations other than those belonging to the M. m. domesticus and M. m. musculus subspecies, implying a possible origin for t-haplotypes prior to the radiation of the most recent offshoot of the Mus genus (i.e., the spretus/domesticus divergence), some 1-3 Myr ago.      

The contractile basis of amoeboid movement: V. The control of gelation, solation, and contraction in extracts from dictyostelium discoideum

Motile extracts have been prepared from Dictyostelium discoideum by homogenization and differential centrifugation at 4 degrees C in a stabilization solution (60). These extracts gelled on warming to 25 degrees Celsius and contracted in response to micromolar Ca++ or a pH in excess of 7.0. Optimal gelation occurred in a solution containing 2.5 mM ethylene glycol-bis (β-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA), 2.5 mM piperazine-N-N'-bis [2-ethane sulfonic acid] (PIPES), 1 mM MgC1(2), 1 mM ATP, and 20 mM KCI at ph 7.0 (relaxation solution), while micromolar levels of Ca++ inhibited gelation. Conditions that solated the gel elicited contraction of extracts containing myosin. This was true regardless of whether chemical (micromolar Ca++, pH >7.0, cytochalasin B, elevated concentrations of KCI, MgC1(2), and sucrose) or physical (pressure, mechanical stress, and cold) means were used to induce solation. Myosin was definitely required for contraction. During Ca++-or pH-elicited contraction: (a) actin, myosin, and a 95,000-dalton polypeptide were concentrated in the contracted extract; (b) the gelation activity was recovered in the material sqeezed out the contracting extract;(c) electron microscopy demonstrated that the number of free, recognizable F-actin filaments increased; (d) the actomyosin MgATPase activity was stimulated by 4- to 10-fold. In the absense of myosin the Dictyostelium extract did not contract, while gelation proceeded normally. During solation of the gel in the absense of myosin: (a) electron microscopy demonstrated that the number of free, recognizable F- actin filaments increased; (b) solation-dependent contraction of the extract and the Ca++-stimulated MgATPase activity were reconstituted by adding puried Dictyostelium myosin. Actin purified from the Dictyostelium extract did not gel (at 2 mg/ml), while low concentrations of actin (0.7-2 mg/ml) that contained several contaminating components underwent rapid Ca++ regulated gelation. These results indicated : (a) gelation in Dictyostelium extracts involves a specific Ca++-sensitive interaction between actin and several other components; (b) myosin is an absolute requirement for contraction of the extract; (c) actin-myosin interactions capable of producing force for movement are prevented in the gel, while solation of the gel by either physical or chemical means results in the release of F-actin capable of interaction with myosin and subsequent contraction. The effectiveness of physical agents in producting contraction suggests that the regulation of contraction by the gel is structural in nature.     

停乳链球菌isp基因的克隆与原核表达

根据GenBank报道的停乳链球菌(Streptococcus dysgalactiae)isp基因(GenBank:CP002215.1)序列设计引物,PCR扩增得到isp基因。片段大小为1 500 bp,与从GenBank下载的序列进行同源性比对结果为98.57%。将isp基因克隆于表达载体pET-25b,成功构建了重组质粒pET-25b-isp,将重组质粒分别转化BL21(DE3),将BL21(pET-25b-isp)阳性重组菌株经IPTG诱导后,SDS-PAGE结果显示均有蛋白表达,大小分别为53 ku与目的蛋白大小相符;Western blot检测显示,表达的蛋白为目的蛋白。对成功构建的菌株BL21(pET-25b-isp)表达条件经优化后,在温度37℃,诱导时间12 h,IPTG浓度为1 mmol/L时,蛋白有较高的表达量。     

生产力调控对翌年苹果园土壤水分和苹果叶片光合特性的影响

以盛果期‘长枝富士’为试验材料, 以生产力调控翌年苹果(Malus pumila)树坐果期为主要观测期, 对不同生产力水平下果园土壤水分状况及苹果叶片光合特性进行了研究。结果表明: 在生产力调控范围内, 土壤剖面各个层次土壤含水量均随着生产力水平的减小而增加, 其中在60 cm处达到最大增量, 为31.02%; 而在600 cm范围内土壤贮水量最大能够提高15.41%。随着生产力水平的降低, “光合午休”现象减弱, 净光合速率(P_n)增加, 最大增幅为25.71%, 下午时段的蒸腾速率(T_r)下降迅速, 水分利用效率(WUE)最大提高34.12%。通过相关分析表明, 土壤贮水量(WSC)与P_n、T_r、WUE之间均达到显著相关, 其相关系数分别为: 0.973^**、-0.543^*和0.992^**。土壤贮水量(x)与水分利用效率(y)之间符合y = 0.002 3x - 1.480 6, R²= 0.984 4^**的回归模型。通过生产力调控可以改善土壤水分状况和果树光合能力, 提高WUE。