一种纯化细菌有机磷水解酶的简便方法

Cibacron blue T_3GA与溴化氰活化的Sepharose 4B偶联后,产生一种能有效地分离有机磷水解酶的吸附剂。用0.15mol/L MgCl_2溶液从黄杆菌P3—2细胞抽提出的粗酶液通过柱层析分离,即可得到纯化8倍、酶活性回收率为269.4％的纯酶制品。该酶制品用凝胶电泳测是均一的。     

Application of automated DNA sizing technology for genotyping microsatellite loci.

Highly polymorphic microsatellite loci offer great promise for gene mapping studies, but fulfillment of this potential will require substantial improvements in methods for accurate and efficient genotyping. Here, we report a genotyping method based on fluorescently labeled PCR primers and size characterization of PCR products using an automated DNA fragment analyzer. We capitalize on the availability of three distinct fluorescent dyes to label uniquely loci that overlap in size, and this innovation increases by threefold the number of loci that can be analyzed simultaneously. We label size standards with a fourth dye and combine these with the microsatellite PCR products in each gel lane. Computer programs provide very rapid and accurate sizing of microsatellite alleles and efficient data management. In addition, fluorescence signals are linear over a much greater range of intensity than conventional autoradiography. This facilitates multiplexing of loci (since signal intensities often vary greatly) and helps distinguish major peaks from artifacts, thereby improving genotyping accuracy.     

Species-specific diversity among simian immunodeficiency viruses from African green monkeys

The prevalence, natural history, and genetic characteristics of simian immunodeficiency virus (SIV) infections in most feral African monkey species are presently unknown, yet this information is essential to elucidate their origin and relationship to other simian and human immunodeficiency viruses. In this study, a combination of classical and molecular approaches were used to identify and characterize SIV isolates from West African green monkeys (Cercopithecus sabaeus) (SIVagm isolates). Four SIVagm viruses from wild-caught West African green monkeys were isolated and analyzed biologically and molecularly. Amplification, cloning, and sequencing of a 279-bp polymerase fragment directly from uncultured peripheral blood mononuclear cells was facilitated by the use of nested polymerase chain reaction. The results indicated that West African green monkeys are naturally infected with SIVs which are closely related to East African SIVagm isolates. However, structural, antigenic, and genetic differences were observed which strongly suggest that the West African green monkey viruses comprise a phylogenetically distinct subgroup of SIVagm. These findings support our previous hypothesis that SIVagm viruses may have evolved and diverged coincident with the evolution and divergence of their African green monkey host. In addition, this study describes a polymerase chain reaction-based approach that allows the identification and molecular analysis of divergent SIV strains directly from primary monkey tissue. This approach, which does not depend on virus isolation methods, should facilitate future studies aimed at elucidating the origins and natural history of SIVs in feral African green monkey populations.     

A conserved coronavirus epitope, critical in virus neutralization, mimicked by internal-image monoclonal anti-idiotypic antibodies.

Monoclonal antibody (MAb) 6A.C3 neutralizes transmissible gastroenteritis coronavirus (TGEV) and is specific for a conserved epitope within subsite Ac of the spike (S) glycoprotein of TGEV. Six hybridomas secreting anti-idiotypic (Ab2) MAbs specific for MAb 6A.C3 (Ab1) have been selected. All six MAbs inhibited the binding of Ab1 to TGEV and specifically cross-linked MAb1-6A.C3. Four of these hybridomas secreted gamma-type anti-idiotypic MAbs. The other two Ab2s (MAbs 9A.G3 and 9C.E11) were recognized by TGEV-specific antiserum induced in two species. This binding was inhibited by viruses of the TGEV group but not by serologically unrelated coronaviruses. These results indicate that MAb2-9A.G3 and MAb2-9C.E11 mimic an antigenic determinant present on the TGEV surface, and they were classified as beta-type ("internal-image") MAbs. TGEV-binding Ab3 antiserum was induced in 100% of mice immunized with the two beta-type MAb2s and in 25 to 50% of mice immunized with gamma-type MAb2. Both beta- and gamma-type Ab2s induced neutralizing Ab3 antibodies in mice that were mainly directed to antigenic subsite Ac of the S protein.     

Carbon-starvation induction of the ugp operon, encoding the binding protein-dependent sn-glycerol-3-phosphate transport system in Escherichia coli.

Summary The gene products of the ugp operon of Escherichia coli are responsible for the uptake of sn-glycerol-3-phosphate and certain glycerophosphodiesters. The regulation of ugp is mainly phoBR-dependent. Significant expression, however, can be observed even in the presence of high concentrations of phosphate, a condition which normally completely represses pho expression. Pho-independent ugp expression was found to be derepressed during the late logarithmic growth phase due to carbon starvation. Among different carbon sources tested, glucose caused the most complete repression. Addition of cAMP prevented glucose repression, indicating that a cAMP-CRP control mechanism may be directly or indirectly involved in the carbon-starvation response. This conclusion is supported by the fact that pho-independent ugp expression correlated with the presence of the cya and crp gene products.     

The reconciliation behavior of golden monkeys (Rhinopithecus roxellanae roxellanae) in small breeding groups

Observations were made following 130 spontaneous aggressive incidents in two small breeding groups of captive golden monkeys (Rhinopithecus roxellanae roxellanae). Participants were observed both during the first 10 min following these incidents and during matched control observations. An increased contact rate was observed between opponents following the aggressive incident. Post-conflict contacts were characterized by a number of behavior patterns: open mouth, rapid grooming, embrace, and crouching. Adult males played an important role as mediator in agonistic disputes among females: males intervened in 93.6% of female fights. It is speculated that this intervention behavior is related to the species' organization into one-male units.     

湘中、湘东地区早籼稻耐土壤潜育性评价

我国南方稻区的主要低产稻田是潜育性稻田,约有一亿亩。挖掘其“潜在生产力”,种植耐潜育性土壤逆境胁迫能力较强的水稻品种,则是简便、经济而有效的重要途径之一。本文就几个早籼稻品种(组合)对潜育性稻田的生态适应性进行了较系统的观测,并初步提出了耐潜育性的几个鉴定指标,诸如根系生长量和幼穗分化期根系氧化力;分蘖早期茎蘖增长速率;分蘖后期单株干物质产量;乳熟期剑叶片过氧化氢酶活性GDI和光合强度等。上述鉴定指标,综合应用于水稻品种生态适应性和耐潜育性育种研究,有助于提高水稻抗逆性育种的效率。     

慈竹无性系种群生长发育规律初探

慈竹(Neosinocalamus affinis)系四川分布面积最大、产量高、用途广的重要经济竹种,对振兴四川经济具有很大的潜在作用。作者引用Harper的构件结构理论,将慈竹无性系作为一个种群进行研究。以慈竹幼竹为研究对象,通过对其生长发     

Molecular cloning and expression of the human deoxythymidylate kinase gene in yeast.

(Deoxy)thymidylate (dTMP) kinase is an enzyme which phosphorylates dTMP to dTDP in the presence of ATP and magnesium. This enzyme is important in cellular DNA synthesis because the synthesis of dTTP, either via the de novo pathway or through the exogenous supply of thymidine, requires the activity of this enzyme. It has been suggested that the activities of the enzymes involved in DNA precursor biosynthesis, such as thymidine kinase, thymidylate synthase, thymidylate kinase, and dihydrofolate reductase, are subjected to cell cycle regulation. Here we describe the cloning of a human dTMP kinase cDNA by functional complementation of a yeast dTMP kinase temperature-sensitive mutant at the non-permissive temperature. The nucleotide sequence of the cloned human cDNA is predicted to encode a 24 KD protein that shows considerable homology with the yeast and vaccinia virus dTMP kinase enzymes. The human enzyme activity has been investigated by expressing it in yeast. In this work, we demonstrate that the cloned human cDNA, when expressed in yeast, produces dTMP kinase activity.     

The formation and characteristics of China's existing system

Conclusion To sum up, the character of China's existing system is basically all-embracing totalitarianism formed by the combination of the old tradition of feudal-despotism and the new tradition of Stalinism. This system is the basic source of all the social evils in China.The ultimate aim of the democratic movement is to transform the old, allembracing totalitarian system and to establish a democratic, free, humane, and rational new system. The present ruling group is the beneficiary and defender of the existing system. The contradictions and conflicts between reformers who want to transform the old system and the hardliners who want to protect the old system formed the crux of the struggle of the 1989 democratic movement. Because the democratic movement lacked preparation in theory, organization, tactics and strategy, and because the ruling group controls the means of dictatorship — especially the army —military force was used to suppress the democratic movement with brutal bloodshed. In order to promote the democratic movement, we should sum up some lessons and experiences. First, we should study in depth the existing system and oppose feudaldespotism and Stalinism through effective propaganda and education. Second, we should arouse the democratic consciousness of the broad masses of the people, uniting with them and the healthy elements inside the Party, government, and army to struggle for the smashing of the old system and the establishment of a new system.Su Shaozhi is former Director of the Institute for the Study of Marxism-Leninism and Mao Zedong Thought, Chinese Academy of Social Sciences, Beijing, and currently Visiting Professor at the Bradley Institute for Democracy and Public Values, Marquette University, Milwaukee, Wisconsin