Deep-level diagnostic value of the rDNA-ITS region

The similarity of certain reported angiosperm rDNA internal transcribed spacer (ITS) region sequences to those of green algae prompted our analysis of the deep-level phylogenetic signal in the highly conserved but short 5.8S and hypervariable ITS2 sequences. We found that 5.8S sequences yield phylogenetic trees similar to but less well supported than those generated by a ca. 10-fold longer alignment from rDNA-18S sequences, as well as independent evidence. We attribute this result to our finding that, compared to 18S, the 5.8S has a higher proportion of sites subject to vary and greater among-site substitution rate homogeneity. We also determined that our phylogenetic results are not likely affected by intramolecular compensatory mutation to maintain RNA secondary structure nor by evident systematic biases in base composition. Despite historical homology, there appears to be no ITS2 primary sequence similarity shared sufficient similarity to cluster correctly on the basis of alignability. Our results indicate that groups, however, share sufficient similarity to cluster correctly on the basis of alignability. Our results indicate that ITS region sequences can diagnose organismal origins and phylogenetic relationships at many phylogenetic levels and provide a useful paradigm for molecular evolutionary study.      

Kinetics of retroviral production from the amphotropic ΨCRIP murine producer cell line

Rapidly expanding development and practice of gene therapy requires the availability of large quantities of high titer retroviral supernatants. One way to achieve high retroviral titers is through improved understanding of the kinetics of retroviral production and decay, and the subsequent development of improved cell culture methods. In the present study we investigated the effects of different operational modes on the retroviral production of the NIH 3T3 fibroblast derived amphotropic murine retroviral producing cell line pMFG/CRIP. Semi-continuous culture (exchange of 50% of medium volume daily) was found to promote cell growth and enhance retroviral production. The rapid medium exchange resulted in significantly larger amounts of high titer supernatants and an extended production phase as compared to the batch control cultures. The specific viral productivity of the pMFG/CRIP cells was in the range of 10 to 40 infectious viruses produced per thousand producer cells per day. The CV-1 African Green Monkey kidney cell line was used as the infection target. Lowering the serum level form 20% to 10% improved retroviral production slightly. However, at lower serum levels (1%, 5% and 10% (v/v)) growth of the producer cell line, and thus retroviral production, was directly proportional to the serum level. The half-life of the virus at 37°C was found to be 5.5 hours. Promoting the growth of producer cell lines can improve retroviral vectors titers and viral production. High cell density systems that allow for rapid cell growth and waste product removal are likely to be used to generate high-titer retroviral supernatants.     

Influences on the antimicrobial activity of surface-adsorbed nisin

The efficacy of the antimicrobial peptide nisin was examined after adsorption to silica surfaces. Three protocols were used to evaluate nisin's activity against adhered cells ofListeria monocytogenes: bioassay usingPediococcus pentosaceous FBB 61-2 as the sensitive indicator strain; visualization and enumeration of cells by microscopic image analysis; and viability of adhered cells as determined by lodonitrotetrazolium violet uptake and crystallization. The activity of adsorbed nisin was highly dependent upon conditions of adsorption. The highest antimicrobial activity of adsorbed nisin occurred with high concentrations of nisin (1.0 mg ml^–1) and brief contact times (1 h) on surfaces of low hydrophobicity. Sequential adsorption of a second protein (-lactoglobulin or bovine serum albumin) onto surfaces consistently resulted in decreased nisin activity. These data provide direction for the development of applications to limit microbial attachment on food contact surfaces through the use of adsorbed antimicrobial peptides.     

包囊游仆虫休眠包囊的超微结构研究

包囊游仆虫休眠包囊中,各类纤毛器的纤毛基体上方的大部分纤毛杆退化,或仅保留毛基体,有时部分额腹棘毛的毛基体也瓦解消失。残留纤毛的纤毛杆周围微管和中央微管仍具有“9 2”结构特征,也有少数纤毛杆出现2套“9 2”微管共处于一层纤毛膜内的现象。毛基体中周围三联体微管的中央形成微管形结构聚合体,基体附属结构仅存在基体间连接及纤毛器托架的残余物;非纤毛区皮层表膜下未见微管层。纤毛区皮层含纤毛器腔周围微管层(相当于表膜下微管层)、纤毛器深部及附近的微管束和分散的微管群。并且,纤毛区皮层囊泡内含有呈不同形态的纤毛杆结构;大核核孔明显变大,核孔数目减少,核孔内膜附着染色质。     

山西雏蝗属二新种：直翅目：网翅蝗科

本文记述山西省雁北地区雏蝗属Ｃｈｏｒｔｈｉｐｕｓ Ｆｉｅｂｅｒ１８５２二新种,即郑氏雏蝗Ｃｈｏｒｔｈｉｐｐｕｓ ｚｈｅｎｇｉ,ｓｐ．ｎｏｖ．和宽带雏蝗Ｃｈｏｒｔｈｉｐｐｕｓ ａｍｐｌｉｌｉｎｅａｔｕｓ,ｓｐ．ｎｏｖ。模式标本保存在山西省农牧厅植保植检总结。     

简便、快速提取高质量的RNA分子

MethodsforHandy,RapidIsolationofHighQualityRNAbyGuanadiumThioeyauateDuJianYuanYanhuaMaShenglinDongZhiwei(BeijingInstituteforCancerResearchBeijing100034)硫氰胍是一种有效的蛋白变性剂。早在1979年,Chirgwin等就利用氯化铯／硫氰胍超离心技术成功地从RNA酶富集的胰脏组织中提取出未降解的RNA分子[2],从而使它成为抑制RNA酶的首选药物并得到广泛使用,但受到超速离心设备的限制。1983年,Cathala报道了氯化锂／硫氰胍RNA提取法[1]。该方法操作简便,获得的RNA质量很高,但所需时间较长。为了能在短时间内更快…     

c-fos expression in the amygdala:In vivo antisense modulation and role in anxiety

Summary 1. The amygdaloid complex is a key structure in mechanisms of fear and anxiety. Expression of the immediate-early gene c-fos has been reported in the central nucleus of the amygdala following various stressors, but the functional role of this phenomenon has remained unknown.2. c-fos expression was observed in the central nucleus when rats were subjected to a pharmacologically validated animal model of anxiety, the Vogel conflict test, but not after mere exposure to the test apparatus. Bilateral amygdala injection of a 15-mer phosphorothioate c-fos antisense oligodeoxynucleotide prior to testing blocked conflict-induced c-fos expression and had behavioral effects similar to those of established antianxiety drugs.3. Separate experiments determined that antisense treatment did not affect conflict behavior by acting on shock thresholds or drinking motivation.4. These findings provide evidence that neuronal activation and c-fos induction in the amygdala may be of importance for mechanisms of fear and anxiety.     

Multiple genotypes of mitochondrial DNA within a horse population from a small region in Yunnan province of China

mtDNA genotypes of six domestic horses (three adult short horses whose heights are under 1 m and three common domestic horses) from a small region of 15 km² in Malipo county of Yunnan province of China were investigated by the technique of restriction fragment length polymorphism (RFLP) with 16 restriction endonucleases which recognize 6-bp sequences. An average of 56 fragments for an individual was obtained. Unlike other domestic animals, this population of horses exhibits high mtDNA genetic diversity. Each of the six horses has a specific mtDNA genotype showing a pattern of multiple maternal origins, as suggested by fossil and literature records. We think the population of horses is an amazing seed-resource pool of horses and hence deserves to be paid more attention from the view of conservation genetics. However, it is also remarkable that we did not find any typical mtDNA genetic markers which would discriminate between short horses and common domestic horses.     

Comparative analysis of multiple protein-sequence alignment methods [published erratum appears in Mol Biol Evol 1994 Sep;11(5):811]

We have analyzed a total of 12 different global and local multiple protein-sequence alignment methods. The purpose of this study is to evaluate each method's ability to correctly identify the ordered series of motifs found among all members of a given protein family. Four phylogenetically distributed sets of sequences from the hemoglobin, kinase, aspartic acid protease, and ribonuclease H protein families were used to test the methods. The performance of all 12 methods was affected by (1) the number of sequences in the test sets, (2) the degree of similarity among the sequences, and (3) the number of indels required to produce a multiple alignment. Global methods generally performed better than local methods in the detection of motif patterns.      

菜用香椿良种选育性状的模糊综合评判

采用模糊数学方法对菜用香棒１６个不同种源的无性系从形态、颜色、香气、抗病性四个方面进行了综合评判,筛选出了接近选育目标的５个菜用优良无性系。