羊传染性脓疱病毒新疆流行株F1L基因克隆及表达

目的:为了探讨F1L重组蛋白作为羊传染性脓疱病毒亚单位疫苗的可行性。方法:以分离出羊传染性脓疱病毒ORFV-shz分离株为模板,扩增F1L基因片段,并将其克隆至pMD18-T载体中进行测序。构建原核表达载体pET-32a-F1L,转化至大肠埃希菌BL21,用SDS-PAGE及Western blot检测目的蛋白的表达情况及其反应原性。结果:PCR扩增出F1L基因全长1023bp,共编码340个氨基酸;推导的氨基酸序列中第1～70位氨基酸构成信号肽序列,第285～313位氨基酸为跨膜区。SDS-PAGE分析表明获得了约57.4kDa的融合蛋白,在Western-blotting检测中,融合蛋白可与羊传染性脓疱病毒阳性血清发生特异性反应,表明其具有良好的反应原性。结论:成功构建F1L基因的原核表达载体,为研究新疆地区羊传染性脓疱的防治及疫苗开发提供了科学依据。     

Aspochracin,a New Insecticidal Metabolite of Aspergillus ochraceus

A new insecticidal substance named aspochracin was isolated from the culture filtrate of Aspergillus ochraceus, a pathogenic fungus causing muscardine on insects. The compound was found to be a novel cyclotripeptide, composed of N-methyl-l-alanine, N-methyl-l-valine and l-ornithine, containing an octatrienoic acid side chain. Isolation procedure, structural elucidation and biological activities are described in detail.

Hexahydroaspochracin (II) derived on hydrogenation of aspochracin (I), an insecticidal metabolite of Aspergillus ochraceus, was synthesized by cyclization of N-methyl-l-valyl-N- methyl-l-alanyl-α-caprylyl-l-ornithine (XIV). In addition to II, isohexahydroaspochracin (XV) was isolated from the reaction mixture.     

OB增效剂对斜纹夜蛾核多角体病毒的增效作用

本文以OB作为斜纹夜蛾核多角体病毒（Spodoptera litura nucleopolyhedrovirus,SpltNPV）的增效剂,对斜纹夜蛾Spodoptera litura幼虫进行生物测定,结果表明,在0．25％-1．00％的浓度范围内,随着OB增效剂浓度的提高,其对斜纹夜蛾核多角体病毒的增效作用也随着提高,最高增效倍数达85．1倍;在2～4龄幼虫范围内,随着虫龄的增大,OB增效剂对斜纹夜蛾核多角体病毒的增效作用也增加;而随着温度的升高,增效剂的增效作用无显著提高。     

Estimating the intensity of male-driven evolution in rodents by using X-linked and Y-linked Ube 1 genes and pseudogenes

Using sequence data from the last introns of ZFX and ZFY genes, we previously estimated the male-to-female ratio () of mutation rate to be close to 6 in higher primates and 1.8 in rodents. As the mutation rate may vary among different regions of the mammalian genome, it is interesting to see whether sequence data from other regions will give similar estimates. In this study, we have determined the partial genomic sequences of the ubiquitin-activating enzyme El genes (Ube 1x and Ube 1y for the X-linked and Y-linked homologues, respectively) of mice and rats and two mouse Ube 1y pseudogenes. From the intron sequences of the Ube 1 genes, we calculated the divergence of the Y-linked genes (Y = 0.161) and that of the X-linked genes (X = 0.107) between mouse and rat, and found the Y/X ratio to be 1.50. This ratio led to an estimate of = 2.0 with a 95% confidence interval of (1.0, 3.9). Similar estimates of were obtained if mouse Ube 1y pseudogenes were used instead of the mouse Ube 1y functional gene. These estimates are consistent with our previous estimate for rodents and suggest that the sex ratio of mutation rate in rodents is approximately only one-third of that in higher primates. Our estimate of the divergence time between Ube 1x and Ube 1y supports the view that the two genes separated before the eutherian radiation.Correspondence to: W.-H. Li     

Colorimetric determination of urinary adenosine using aptamer-modified gold nanoparticles

This paper describes a colorimetric sensing approach for the determination of adenosine triphosphate (ATP) using aptamer-modified gold nanoparticles (Apt-Au NPs). In the absence of the analytes, the color of the Apt-Au NPs solution changed from wine-red to purple as a result of salt-induced aggregation. Binding of the analytes to the Apt-Au NPs induced folding of the aptamers on the Au NP surfaces into four-stranded tetraplex structures (G-quartet) and/or an increase in charge density. As a result, the Apt-Au NPs solution was wine-red in color in the presence of the analytes under high salt conditions. For mixtures of ATP (20.0–100.0 nM), Apt-Au NPs (3.0 nM), 10.0% poly(ethylene glycol), 0.2 μM TOTO-3, 150.0 mM NaCl, 15.0 mM KCl, and 16.0 mM Tris–HCl (pH 7.4), a linear correlation (R² = 0.99) existed between the ratio of the extinctions of the Apt-Au NPs at 650 and 520 nm (Ex_650/520) and the concentration of ATP. The limit of detection for ATP was 10.0 nM. The practicality of this simple, sensitive, specific, and cost-effective approach was demonstrated through the determination of the concentration of adenosine in urine samples.     

Direct electrochemical sensor for label-free DNA detection based on zero current potentiometry

A direct electrochemical DNA biosensor based on zero current potentiometry was fabricated by immobilization of ssDNA onto gold nanoparticles (AuNPs) coated pencil graphite electrode (PGE). One ssDNA/AuNPs/PGE was connected in series between clips of working and counter electrodes of a potentiostat, and then immersed into the solution together with a reference electrode, establishing a novel DNA biosensor for specific DNA detection. The variation of zero current potential difference (ΔE(zcp)) before and after hybridization of the self-assembled probe DNA with the target DNA was used as a signal to characterize and quantify the target DNA sequence. The whole DNA biosensor fabrication process was characterized by cyclic voltammetry and electrochemical impedance spectroscopy with the use of ferricyanide as an electrochemical redox indicator. Under the optimized conditions, ΔE(zcp) was linear with the concentrations of the complementary target DNA in the range from 10nM to 1μM, with a detection limit of 6.9nM. The DNA biosensor showed a good reproducibility and selectivity. Prepared DNA biosensor is facile and sensitive, and it eliminates the need of using exogenous reagents to monitor the oligonucleotides hybridization.     

逆转录病毒载体介导胸苷磷酸化酶在胰腺癌细胞表达

人胸腺嘧啶核苷磷酸化酶（ＴＰ）在一些肿瘤组织中活性增高,但其功能目前了解尚少。构建了表达ＴＰ的重组逆转录病毒载体,直接导入人胰腺癌ＰＣ－２细胞,ｍＰＣＲ扩增、Ｓｏｕｔｈｅｒｎ及Ｎｏｒｔｈｅｒｎ印迹和原位杂交证实转染细胞有外源ＴＰ的整合及表达,酶活性检测发现含外源ＴＰ细胞ＴＰ活性比ＰＣ－２细胞的内源性ＴＰ活性高ＴＰ活性高４０－７０倍,生长曲线和＾３Ｈ－ＴｄＲ参入率检测未发现含外源ＴＰ细胞生物学行为的     

Phenotypic and genetic differences between opaque and translucent colonies of Vibrio alginolyticus

Many pathogens undergo phase variation between rugose and smooth colony morphology or between opaque and translucent colony morphology, which is mainly due to the variation in the surface polysaccharides. In this study, Vibrio alginolyticus ZJ-51 displayed phase variation between opaque, rugose colonies (Op) and translucent, smooth colonies (Tr). Unlike the vibrios reported previously, Tr cells of ZJ-51 enhanced biofilm formation and motility, but they did not differ from Op cells in the quantity of surface polysaccharides produced. Real time PCR was used to analyze the expression of the genes involved in polysaccharide biosynthesis, flagellar synthesis, and the AI-2 quorum-sensing system. The results revealed that the K-antigen capsule gene cluster (which consists of homologs to the cpsA-K in Vibrio parahaemolyticus) and O-antigen polysaccharide gene cluster (which contains homologs to the wza-wzb-wzc) were significantly more transcribed in Tr cells. The AI-2 quorum-sensing genes showed enhanced expression in the Tr variant which also exhibited greater expression of genes associated with polar flagellar biosynthesis. These results suggest that colony phase variation might affect the virulence and survival ability in the stressful environment inhabited by V. alginolyticus.