Sub-lethal effects of deltamethrin residues on the within-crop behaviour and distribution ofCoccinella septempunctata

The behaviour and distribution of adultCoccinella septempunctata L. (Coleoptera: Coccinellidae) were recorded in two plots of winter wheat infested with the cereal aphidsSitobion avenae (F.) andMetopolophium dirhodum (Walk.) (Homoptera: Aphididae). One plot was sprayed with the pyrethroid insecticide deltamethrin at a rate of 6.25 g a.i./ha and the other was left unsprayed. Single ladybird beetles were released sequentially on the ground at the centre of the sprayed and unsprayed plots and their behaviour and position in the crop canopy were recorded at 30 second intervals for a total of 15 min per beetle. Assessments, with fresh beetles, continued for four days after spray application with a total of eighty ladybird beetles observed. The 15 min period was selected to avoid lethal effects and no ladybird beetles were killed or knocked down as a result of exposure to deltamethrin residues during this period. Significant differences were found between the overall behaviour patterns ofC. septempunctata in the untreated and deltamethrin treated plots up to three days after the spray application. Ladybird beetles exposed to deltamethrin residues were observed to walk and groom significantly more frequently and to rest significantly less frequently than those in the unsprayed plot. Significant differences were also found between the observed distribution of ladybird beetles in the sprayed and unsprayed crop canopies, with higher numbers of observations towards the bottom of the crop canopy and on the ground in the deltamethrin treated plot than in the untreated plot during the first two days after deltamethrin application. Upon the foliage itself, ladybird beetles were observed significantly more frequently on the abaxial leaf surface in the deltamethrin treated crop compared with the untreated crop. The results are discussed in terms of possible evidence for the repellency of deltamethrin toC. septempunctata and also the implications for integrated pest management of changes in predator behaviour and crop distribution resulting from sub-lethal uptake of insecticides.     

Lipofection of Synthetic Oligodeoxyribonucleotide Having a Palindromic Sequence of AACGTT to Murine Splenocytes Enhances Interferon Production and Natural Killer Activity

A synthetic 22-mer oligodeoxyribonucleotide having an AACGTT palindrome, AAC-22, induced interferon (IFN) production and augmented the natural killer (NK) activity in murine splenocytes, whereas its analogue, ACC-22, having an ACCGGT palindrome, did not. The binding of AAC-22 to splenocytes was not different from that of ACC-22. Lipofection of AAC-22 to splenocytes remarkably enhanced IFN production and NK cell activity, whereas that of ACC-22 caused little enhancement. These results strongly suggest that the prerequisite for IFN production is not the binding of AAC-22 to the cell surface receptors, but its penetration into the spleen cells.     

精子作载体的转基因鱼研究

本文报道了以精子为载体将美洲拟蝶抗冻蛋白基因导入罗非鱼卵,构建转基因鱼的方法,此法简单易行。斑点杂文和ＳｏｕｔｈｅｒｎＢｌｏｔ杂交结果表明,外源基因的整合率为１８．１％,与其它方法构建转基因鱼的外源基因整合率相近。     

Metabolic engineering of Clostridium acetobutylicum ATCC 824 for increased solvent production by enhancement of acetone formation enzyme activities using a synthetic acetone operon

The ability to genetically alter the product-formation capabilities of Clostridium acetobutylicum is necessary for continued progress toward industrial production of the solvents butanol and acetone by fermentation. Batch fermentations at pH 4.5, 5.5, or 6.5 were conducted using C. acetobutylicum ATCC 824 (pFNK6). Plasmid pFNK6 contains a synthetic operon (the "ace operon") in which the three homologous acetone-formation genas (adc, ctfA, and ctfB) are transcribed from the adc promoter. The corresponding enzymes (acetoacetate decarboxylase and CoA-transferase) were best expressed in pH 4.5 fermentations. However, the highest levels of solvents were attained at pH 5.5. Relative to the plasmid-free control strain at pH 5.5, ATCC 824 (pFNK6) produced 95%, 37%, and 90% higher final concentrations of acetone, butanol, and ethanol, respectively; a 50% higher yield (g/g) of solvents on glucose; and a 22-fold lower mass of residual carboxylic acids. At all pH values, the acetone-formation enzymes were expressed earlier with ATCC 824 (pFNK6) than in control fermentations, leading to earlier induction of acetone formation. Furthermore, strain ATCC 824 (pFNK6) produced butanol significantly earlier in the fermentation and produced significant levels of solvents at pH 6.5. Only trace levels of solvents were produced by strain ATCC 824 at pH 6.5. Compared with ATCC 824, a plasmid-control strain containing a vector without the ace operon also produced higher levels of solvents [although lower than those of strain ATCC 824 (pFNK6)] and lower levels of acids. Strains containing plasmid-borne derivatives of the ace operon, in which either the acetoacetate decarboxylase or CoA-transferase alone were expressed at elevated levels, produced acids and solvents at levels similar to those of the plasmid-control strain. (c) 1993 John Wiley & Sons, Inc.     

Strong and regulated promoters in the cyanobacterium Anabaena PCC 7120

Abstract The strengths of several promoters were assessed in the cyanobacterium Anabaena PCC 7120 by fusing them to luxAB , encoding bacterial luciferase. Two promoters, P _tac and P _psbA , with sequences nearly identical to consensus Escherichia coli σ ⁷⁰ promoters, gave as high or higher expression than the strong Anabaena promoter, P _rbc . P _npt , the natural promoter driving expression of the kanamycin-resistance determinant from Tn5, was poorly expressed in Anabaena . The Lac repressor partially repressed expression from P _tac , permitting regulated expression in Anabaena after induction with isopropyl thiogalactoside to a level 4–5-fold higher than without inducer.     

Patterns of Integration and Expression of Retroviral, Non-Replicative Vectors in Avian Embryos: Embryo Developmental Stage and Virus Subgroup Envelope Modulate Tissue-Tropism

We previously demonstrated that Avian Leukemia Viruses (ALV) carrying the v-myc gene specifically induce two types of tumors, cardiomyocytic tumors when the virus is injected before embryonic day 3 (E3), skin tumors when the virus is injected at E3 or E5.

Aiming to elucidate the mechanisms which determine this time-dependent change in target, we infected chick and quail embryos at E3 and E5 with replication-deficient, lacZ gene-carrying, ALV-based viruses produced by a packaging cell line. Three constructs driven by 3 different Long Terminal Repeats (LTRs) were tested and yielded similar results. When the constructs were inoculated at E3 and the lacZ gene product revealed 5 days later, around 70% of the embryos carried lacZ+ clones in the heart, around 50% had positive clones in the skin anywhere on the body, while a few embryos displayed clones in internal organs (liver, stomach, lungs). Immunocytological identification of the heart cell type(s) expressing the virus revealed that the only cells infected were cardiomyocytes. When the constructs were inoculated at E5, no lacZ+ clones appeared in the heart but all were located in the cephalic skin. In order to examine the relationship between viral integration and expression, DNA of different organs or tissues from lacZ stained embryos was analyzed by PCR. A tight correlation between integration and expression in the heart and in the skin was revealed in most cases. In contrast, a significant PCR signal was often detected in the liver or the stomach despite weak or absent expression as revealed by lacZ+ clones.

We then investigated the influence of envelope glycoprotein subgroups on the tropism of these constructs. The lacZ vector driven by RAV-2 LTRs was packaged as subgroups A, B or E viral particles. The A subgroup, used in the part of the study described above, infects both chick and quail while the B and E subgroups are specific for chick or quail respectively. These B and E subgroups induced lacZ+ clones in the heart (after E3 injection) while no clones or only a few were detected in the skin either after E3 or E5 injection. The following conclusions can be drawn: 1) cardiomyocytes are at E3 the major target for integration and expression of ALV-derived viruses in vivo; 2) targets change rapidly with embryonic age; and 3) tissue-specific infections depend on the envelope subgroup, thus presumably on the presence of the cognate receptor. This study clearly indicates that E3 inoculation of ALV-based retroviral vectors is a simple and powerful method to transfer gene sequences into cardiomyocytes and epidermal cells.     

酿酒酵母合成木栓酮及其衍生物的研究现状及展望

木栓酮及其衍生物在植物中普遍存在且种类繁多,具有丰富的生理药理学活性。木栓酮衍生物是以木栓酮为骨架经细胞色素氧化酶P450(cytochromeP450,CYP450)及UDP葡萄糖醛酸转移酶(UDP-glucuronosyltransferase, UGT)修饰而来。植物中天然木栓酮及其衍生物的含量极低,传统的萃取分离和化学合成效率低、能耗高且污染环境,因此,利用酿酒酵母作为宿主菌生产木栓酮及其衍生物是一种高效且环保的策略。本文从增加前体含量、提高酶活性和产物合成的亚细胞定位等方面介绍并展望了木栓酮在酿酒酵母中高效生产的策略,并介绍了目前几种常见的木栓酮衍生物研究现状,从根据碳骨架相似性挖掘CYP450、蛋白质工程改造CYP450和合成代谢基因簇的挖掘等方面展望了木栓酮衍生物的合成途径解析的新思路。     

生物活性物的生物制造：现状、挑战和发展趋势

生物活性物的生物制造是指利用包括细胞、微生物和酶在内的生物系统生产具有生物活性的天然或合成分子的过程。这些分子可用于制药、化妆品、农业和食品工业等领域,对提高生命质量、延长生命长度具有重要意义。在合成生物学和自动化等技术的推动下,生物制造领域迅速发展,为创造新产品和替代传统产品提供了绿色可持续的生产模式,为生物经济的增长、创新作出了重要贡献。本文结合生物活性物研发及生产情况,简要梳理并分析了国内外生物活性物的现有市场和未来发展。生物制造作为一种绿色、可持续的生产方式,将在生物经济发展中持续发挥重要作用。     

中国山芹属一新种的研究

运用多学科综合分类手段,研究了山芹不同产地居群的变异,发现分布于我国东北部的各居群与华东居群不但存在地理分布上的间断,而且在外部形态、果实及叶柄解剖、核型组成和生物学特性等方面也存在显著差异,因而华东的山芹各居群应独立出来完成一新种──华东山芹OstericumhuadongensisZ.H.PanetX.H.L。