Silver stained polyacrylamide gels and fluorescence-based automated capillary electrophoresis for detection of amplified fragment length polymorphism patterns obtained from white-rot fungi in the genus Trametes

Silver stained denaturing polyacrylamide gels (PAGEs) and fluorescent denaturing automated capillary electrophoresis (CE) were used to detect amplified fragment length polymorphism (AFLP) patterns obtained from white-rot fungi belonging to the genus Trametes. AFLP fingerprinting detected by the fluorescence-based method as well as by silver staining showed a high discriminatory power in differentiating nine strains of Trametes ochracea, nine strains of Trametes hirsuta and ten isolates of Trametes versicolor. UPGMA dendrograms derived from fluorescently labelled and silver stained AFLP patterns were similar, but a few differences were detected especially in the clustering of T. ochracea and T. hirsuta strains. Compared to silver-stained AFLP, detection of fluorescent AFLP was fast, reliable and easy to perform and it facilitated surveying with a computerized analysis system. Fluorescent CE seems to be well suited for studying similarity between Trametes species.     

Response of a Pseudomonas aeruginosa biofilm community to DNA-damaging chemical agents

Pseudomonas aeruginosa strain RM4440 carries a plasmid-borne fusion of the P. aeruginosa recA gene promoter to a promoterless lux operon from Vibrio fisheri. We tested the response of RM4440 in a biofilm to exposure to a 1-h pulse of each of 17 chemicals known to be toxic to bacteria and other organisms. The induction of light produced from the recA-lux fusion present in RM4440 proved to be sensitive and specific for DNA-damaging chemicals when included in a biofilm environment. This study demonstrates the potential usefulness of this construct for in situ investigation of bacterial communities.     

Carbofuran degradation mediated by three related plasmid systems

Two carbofuran-metabolizing Sphingomonas strains, TA and CD, were isolated from soils with differing histories of exposure to carbofuran. These strains were compared with a previously described strain, Sphingomonas sp. CFO6, with regard to growth rate, formation of metabolites, and plasmid content and structure. Extensive regions of similarity were observed between the three different plasmid systems as evidenced by cross hybridization. In addition, all three systems harbor IS1412, an insertion sequence (IS) element involved in heat-induced loss of carbofuran phenotype in CFO6, and heat-induced carbofuran deficient mutants of all three strains correlated with loss of IS1412. A carbofuran deficient mutant of TA generated by induction of IS elements was complemented by reintroduction of the wild-type plasmid, confirming the presence of genes required for carbofuran metabolism on this plasmid. Carbofuran metabolism in these three strains is clearly linked via plasmids of different numbers and sizes that share extensive common regions, and carbofuran-degrading genes may be associated with active IS elements.     

Sex-linked genes and linkage maps in amphibians

This paper reviews sex-linked genes and linkage maps in amphibians. It appears that there is no common ancestral or conserved sex-linkage group in amphibians, whereas an important proportion of other linkage groups has been conserved in amphibians. Comparisons of amphibian linkage maps with those of fishes and mammals reveal several syntenic associations apparently conserved over a very long period of vertebrate divergence.     

Cell-to-cell contact results in a selective translocation of maternal human immunodeficiency virus type 1 quasispecies across a trophoblastic barrier by both transcytosis and infection

Mother-to-child transmission can occur in utero, mainly intrapartum and postpartum in case of breastfeeding. In utero transmission is highly restricted and results in selection of viral variant from the mother to the child. We have developed an in vitro system that mimics the interaction between viruses, infected cells present in maternal blood, and the trophoblast, the first barrier protecting the fetus. Trophoblastic BeWo cells were grown as a tight polarized monolayer in a two-chamber system. Cell-free virions applied to the apical pole neither crossed the barrier nor productively infected BeWo cells. In contrast, apical contact with human immunodeficiency virus (HIV)-infected peripheral blood mononuclear cells (PBMCs) resulted in transcytosis of infectious virus across the trophoblastic monolayer and in productive infection correlating with the fusion of HIV-infected PBMCs with trophoblasts. We showed that viral variants are selected during these two steps and that in one case of in utero transmission, the predominant maternal viral variant characterized after transcytosis was phylogenetically indistinguishable from the predominant child's virus. Hence, the first steps of transmission of HIV-1 in utero appear to involve the interaction between HIV type 1-infected cells and the trophoblastic layer, resulting in the passage of infectious HIV by transcytosis and by fusion/infection, both leading to a selection of virus quasispecies.     

日利用l基因工程培育成功耐高温植物

Nature Biotechnology 2000年18卷3期249页报道：日本九州大学的科学家们已培育成功一种可在高温下维持光合作用的转基因烟草(Science287,476～479,2000)。鉴于沙漠植物的叶绿体可在高温下减少三烯脂肪酸的合成,科学家们培育了含有来自拟南芥(Arabidopsis thaliana)的附加的叶绿体特异△-3去饱和酶(脱氢酶)基因的两个转基因烟草株系。互阻遏作用导致△-3去饱和酶基因在这些植株中的表达下降,以及与此伴随的三烯脂肪酸的相应减少。因此,这种转基因植株40℃高温下的光合作用活性可高于25℃下的活性(但40℃可极大地降低野生型植株的光合作用强度)。科学家们提示,利用内源基因可避免利－46用外源DNA进行基因操作时发生的有害效应。这个研究组的植物生理学家KohIba说：“此项研究给予我们的启迪是,利用基因操作技术可育成新的农作物或树木,以适应无可避免的全球性气候变化。例如可使寒冷地区的树木适应于高温环境”。汪开治     

应用基因工程技术生产IL-11和IL-12

目前武汉大学生命科学学院科研人员针对人体失血和免疫功能下降,应用基因工程技术生产出了人白细胞介素-11(即IL-11)和人白细胞介素-12(即IL-12)。 1 IL-11为人体自然产生的细胞生长因子,能增加血小板,提高凝血速度,防止伤口出血不止,在临床上用来治疗低血小板症,还能辅助治疗癌症,主要用于化疗。克隆于载体,通过大肠杆菌充分表达,经分离、纯化和精制,生产出质量很高的医用成药,故疗效很好。 2 IL-12这是目前在细胞素中发现对人体免疫活性细胞诱导和调节最强、范围最广的一种。其研制技术是将IL-12的两种不同来源的基因克隆于昆虫病毒载体,在昆虫细胞内表达,经分离纯化成工程蛋白,再制备成医药产品。它能诱导干扰素的产生,激活T细胞和NK细胞产生肿瘤坏死因子,能抗利什曼原虫、疟原虫、结核杆菌、血吸虫、肿瘤和病毒及其各相应的疾病,故美国基因研究所和惠施-阿耶斯特制药公司将它用于治疗艾滋病和肿瘤病,效果很好。秦春圃