Adherence Patterns and DNA Probe Types of Escherichia coli Isolated from Diarrheal Patients in China

One hundred and seventy-two strains of Escherichia coli isolated from diarrheal patients in Beijing, P. R. China, were analyzed for plasmid DNA profile, HEp-2 cell adherence ability and reactivity to 10 previously described DNA probes. They had not been recognized as pathogenic E. coli in China. Of the 110 strains tested, 76 (69%) contained one or multiple large plasmids. Of the 71 strains with the large plasmids 64 could adhere to HEp-2 cells. Of the 172 strains, 102 (59.3%) were hybridized with at least one of the 10 probes. Of those, seven strains hybridized with enteroaggregative E. coli (EAggEC) probe. Their serotypes were O128 (two strains), O6 (one strain), and O111 (one strain). Three strains were untypable. Six and three strains were hybridized with enteropathogenic E. coli (EPEC) attaching and effacing genes (eae) or EPEC adherence factor (EAF) probe, respectively. Two non-O157: H7 strains hybridized with enterohemorrhagic E. coli (EHEC) probe. Seventy-two strains (41.9%) hybridized with shiga-like toxin 2 or 1 (SLT2 or SLT1) probes. Among the SLT1 or SLT2 probe-positive strains, 54 hybridized with invasive (INV) plasmid probe developed for identification of enteroinvasive E. coli (EIEC) and Shigella species. The INV and SLT probe-positive strains might represent a new variety of verotoxin-producing E. coli (VTEC).     

Actions of Rho family small G proteins and p21-activated protein kinases on mitogen-activated protein kinase family members.

The mitogen-activated protein (MAP) kinases are a family of serine/threonine kinases that are regulated by distinct extracellular stimuli. The currently known members include extracellular signal-regulated protein kinase 1 (ERK1), ERK2, the c-Jun N-terminal kinase/stress-activated protein kinases (JNK/SAPKs), and p38 MAP kinases. We find that overexpression of the Ste20-related enzymes p21-activated kinase 1 (PAK1) and PAK2 in 293 cells is sufficient to activate JNK/SAPK and to a lesser extent p38 MAP kinase but not ERK2. Rat MAP/ERK kinase kinase 1 can stimulate the activity of each of these MAP kinases. Although neither activated Rac nor the PAKs stimulate ERK2 activity, overexpression of either dominant negative Rac2 or the N-terminal regulatory domain of PAK1 inhibits Ras-mediated activation of ERK2, suggesting a permissive role for Rac in the control of the ERK pathway. Furthermore, constitutively active Rac2, Cdc42hs, and RhoA synergize with an activated form of Raf to increase ERK2 activity. These findings reveal a previously unrecognized connection between Rho family small G proteins and the ERK pathway.     

Cellobiose-oxidizing enzyme from a newly isolated cellulolytic bacterium Cytophaga sp. LX-7

Summary The cellobiose oxidizing enzyme of the newly isolated cellulolytic bacterium Cytophaga sp. LX-7 was produced extracellularly when grown on cellulose or other saccharides, which was previously noted only in fungi. The enzyme could use not only cellobiose, maltose, glucose and other saccharides but also cellulose as substrates, and use dichlorophenol indophenol and oxygen as electron acceptors.     

Structure and analysis of phospholipase D gene from Ricinus communis L

Phospholipase D (PLD; EC 3.1.4.4) has been proposed to play a pivotal role in various cellular processes, but molecular understanding of this enzyme is rather limited. This report describes the nucleotide sequence, structure, and genomic organization of a PLD gene from castor bean (Ricinus communis L. cv. Hale). The PLD gene was isolated from a castor bean genomic library using the PLD cDNA as a hybridization probe. Sequence comparison with the PLD cDNA revealed that the PLD gene consisted of four exons and three introns, one of which interrupts the 5-untranslated region. Southern blot analysis indicated that the cloned PLD gene was present as a single-copy gene, and yet there were other PLD or PLD-related sequences in the castor bean genome.     

组织型纤溶酶原激活剂（t—PA）工程细胞系遗传特性及成瘤性分析

组织型纤溶酶原激活剂(t-PA)因其在防止血栓形成中起重要作用而受到人们的重视。但由于t-PA在血液中半衰期很短,作为溶栓药,一时难于推广。为了延长半衰期、增强其特异活性,本组构建了t-PA突变体并在CHO-dhfr~-细胞中获得了高效表达。我们在细胞培养基中加入秋水仙素,通过低张、固定、染色,进行染色体分析,结果表明,t-PA工程细胞系染色体条数为20条,畸变类型有异着丝粒。四倍体、裂隙、断片,畸变率为15％,属于正常范围。同时我们对该细胞系进行成瘤性试验,选用4周龄裸鼠作为试验鼠,以Hela细胞为阳性对照,CHO-dhfr~-细胞为阴性对照,试验表明:t-PA工程细胞及表达产物对裸鼠均无成瘤性。     

Effect of sulphate on photosynthesis in greenhouse–grown tomato plants

Sulphate accumulates in the rhizosphere of plants grown in hydroponic systems. To avoid such sulphate accumulation and promote the use of environmentally sound hydroponic systems, we examined the effects of four sulphate concentrations (0.1, 5,2, 10.4 and 20.8 m M ) on photosynthesis, ribulose-l,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) activities and related physiological processes in greenhouse–grown tomato plants ( Lycopersicon esculentum Mill. cv. Trust). The lowest sulphate concentration (0.1 m M ) significantly decreased photosynthetic capacity (P_c) and Rubisco activities on a leaf area basis. This result was supported by our data for dry matter per plant, which was low for plants in the 0.1 m M treatment. The photosynthesis-related variables such as leaf conductance, chlorophyll and soluble protein were lowest for the 0.1 m M treatment. Both total Rubisco activity and the activated ratio were reduced with this treatment. However, Rubisco activities expressed per g of protein or per g of chlorophyll were not significantly affected. These results suggest that sulphur deficiency depressed P_c– by reducing the amount of both Rubisco and chlorophyll and by causing an inactivation of Rubisco. The ratio of organic sulphur vs organic nitrogen (S/N) in plants of the 0.1 m M treatment was far below the normal values. This low S/N ratio might be accountable for the negative effect of low sulphate on P_c and plant growth. P_c and dry matter were not affected until sulphate concentration in the nutrient solution reached a high level of 20.8 m M .     

In vitro fertilization of in vitro matured pig oocytes: effects of boar and ejaculate fraction

Ejaculates from 3 young boars were collected on 4 occasions as a series of separate 15-ml fractions. The contribution of different fractions of these ejaculates to observed variability in the quality of the semen when used for IVF was then determined. On the basis of sperm concentration, 3 fractions representing the first peak concentration (Fraction 1), the lowest sperm concentration after Fraction 1 (Fraction 2), and the second peak concentration (Fraction 3) were selected for analysis in vitro. Oocyte-cumulus-granulosa cell complexes were obtained by dissection from slaughterhouse ovaries. In vitro matured oocytes were randomly assigned for fertilization by the 3 semen samples from each boar. Sperm concentration was the same in all the samples during prefertilization incubation, while the final concentration for fertilization was 5 x 10(5) sperm/ml. Data were analysed using ANOVA for a split-plot design. In the presence of fraction effects, Student-Newman-Keuls (SNK) test was used for multiple comparison of treatment means. Oocyte penetration rates differed among fractions (P = 0.001) and varied from 69 to 100% (mean 95.7%) for Fraction 1, from 0 to 100% (mean 53.3%) for Fraction 2, and from 50to 100% (mean 89.9%) for Fraction 3. There were also differences in male pronuclear formation rate (P = 0.028; mean 27.6, 9.3 and 16.4% for Fractions 1, 2 and 3, respectively); in the rate of polyspermy (P = 0.0001; mean 92.3, 31.9 and 76.3% for Fractions 1, 2 and 3, respectively); and in the number of penetrated spermatozoa per oocyte P = 0.002; mean 5.58, 1.94 and 4.07 for Fractions 1, 2, and 3, respectively). The first peak concentration of semen (Fraction 1) showed superiority in fertilizing ability and less variability in penetration rate from replicate to replicate compared with the other 2 fractions. By multiple comparison, Boar 1 showed higher rates of penetration (P < 0.05), male pronuclear formation (P < 0.05) and polyspermy (P < 0.05) than the other 2 boars. There was no fraction-by-boar interaction. The IVM-IVF system adopted proved to be a promising method for boar semen evaluation.     

生物复苏——大绝灭后生物演化历史的第一幕

生命史是一部生物界短期,快速剧变与长期,慢速稳定相互交替的历史。大绝灭（即集群绝灭）事件反映了全球环境的大突变,点断了地质历史中的生命记录及其发展历程,预示着生物界的演化出现了最有意义的飞跃,近年来尝试研究大绝灭后全球生物界的残存－复苏及其基本型式,并探索复苏的控制因素,标志着地质科学中一个重心的转移（即从大绝灭转向其后的生物残存与复苏的研究）。生物复苏揭示了大绝灭后生物演化历史的第一幕,其研究的     

部分酶解酵母高效电击转化研究

以酵母质粒YCp50为外源DNA,电击转化部分酶解酵母宿主菌AB1380,转化效率稳定在10~6转化子/μg质粒DNA左右,比不酶解酵母或酵母原生质球作受体的电击转化效率高一个数量级以上,也比PEG介导的酵母原生质球转化高3～5倍,而且适合于大片段DNA如水稻YAC分子的转化。达最佳转化时的有关技术参数为:新接菌种通气培养至细胞密度1×10~8～1.5×10~8个/ml;转化时细胞密度控制在1×10~9～1.5×10~9个/ml;每毫升酶解缓冲液加15u溶菌酶(lyticase),30℃下处理酵母5min进行部分酶解;电击时,电场设置在6.25kV/cm、电容25μF,电击后直接铺板。