Detection of a protein encoded by a class II mitochondrial intron of Podospora anserina

Summary In the filamentous fungus Podospora anserina, the amplification as circular DNA molecules of the first intron (intron ) of the CO1 mitochondrial gene, encoding the cytochrome oxidase subunit 1, is known to be strongly associated with aging of strains. In this study we have attempted to detect the protein potentially encoded by the open reading frame (ORF) contained in this intron. This was done by the Western blot technique using specific antisera raised against three polypeptides encoded by three non-overlapping fragments of this ORF adapted to the universal code and overexpressed in Escherichia coli. We examined about thirty independent subclones of Podospora derived from two different geographic races (A, s), using wild-type and mutant strains, young and senescent cultures. A 100 kDa polypeptide, encoded by the class II intron , was detected in five senescent subclones which all showed strong amplification of the intronic sequence (Sen DNA ).     

Functional analysis of a complex oncogene arrangement in biotype III Agrobacterium tumefaciens strains

The ubiquitous grapevine-associated octopine/cucumopine Ti plasmids of biotype III Agrobacterium tumefaciens strains carry two T regions, TA and TB, with a complex oncogene arrangement. Within the octopine/cucumopine group, two main strain types were identified: large TA strains with a TA region resembling the TL region of the biotype I octopine strain Ach5 and small TA strains with a similar T region organization as the large TA strains but with a large internal TA deletion. Structural and functional studies of the representative large TA strain Tm4 revealed six oncogenes. Each oncogene was inserted in a disarmed vector and tested for biological activity using the corresponding oncogenes of Ach5 as standards. Five Tm4 oncogenes, TA-iaaM, T-ipt, T-6b, TB-iaaH and TB-iaaM, were shown to be active, the IS-interrupted TA-iaaH gene was inactive. To study the role of each gene in the pTiTm4 context, several single and multiple pTiTm4 mutations were constructed. It was shown that whereas TA-iaaM and TB-iaaH are essential for tumour formation on grapevine, T-ipt, T-6b and TB-iaaM are not. The avirulence of the TA-iaaM ^- mutant was shown to be due to an inhibitory effect of the T-ipt gene, since a TA-iaaM ^- /T-ipt ^- double mutant was fully virulent. We conclude that the TA-iaaM gene of large TA strains is specifically required to counteract the tumour growth inhibiting activity of the T-ipt gene. Both TA-iaaM and T-ipt are absent from the small TA strains. A model on the roles and interactions of the different oncogenes in large TA and small TA strains is presented.     

Single crystals of V amylose complexed with isopropanol and acetone

Single crystals of amylose complexed with isopropanol or acetone were prepared by adding these precipitants to a metastable aqueous solution of amylose. With both precipitants, similar micrometre sized platelet crystals were obtained. They gave indistinguishable electron diffraction diagrams which could be indexed in an orthorhombic unit cell, with a = 28.26 A, b = 29.30 A, c = 8.01 A and in a space group P2(1)2(1)2(1) or P2(1)2(1)2. Within the unit cell, the amylose chains are organized in antiparallel pairs of parallel 6(5) amylose helices occupying 70% of the cell content, the remaining 30% consisting of isopropanol/acetone and water, with an estimate of 10 isopropanol/acetone molecules for 52 water molecules per unit cell. If the crystals are suspended in pure isopropanol at various temperatures or in pure methanol at room temperature, they undergo a de-solvation process that ultimately converts them into VH amylose. De-solvation with isopropanol left the crystals intact whereas with methanol, they became cracked during the shrinkage. An explanation is proposed for such difference.     

Culture analysis and external interaction models of mycelial growth

To investigate morphogenesis and in particular circularization mechanisms in young mycelia, we observe cultures of the zygomyceteMucor spinosus and develop discrete models of two-dimensional filamental branching growth. The models are based on the hypothesis that the fungus secretes a regulatory substance that diffuses into the surrounding medium and is detected by the growing hyphae. We also present a simple Markovian growth model without such a feedback, but yielding to analytical computations.     

Gene technology-based antimetabolite design: the use of an in vitro protein expression system to facilitate antimetabolite design for virally-induced human diseases and malignant conditions

A precondition for the chemotherapeutic treatment of a variety of virally-induced human diseases and malignant conditions is a highly selective interaction of the drug molecule to be used with it's biological target. To ensure the development of novel, effective drugs, it is essential that the biological target is well characterised with regard to it's structure and activity. Such characterisation relies upon adequate amounts of pure target being available. One of the most important enzymatic importers for antimetabolites is the enzyme thymidine kinase. In this article an in vitro protein expression system is described which facilitates the production of milligram amounts of pure and biologically active thymidine kinase, from a number of important biological sources. Results have shown that the in vitro produced enzyme has the exact biochemical propeties of the in vivo enzyme. Thus the in vitro protein expression system is an ideal vechicle to facilitate an in depth investigation of the enzyme's biological properties.     

中国食蚜蝇科—新属新种(英文)

本文记述了中国食蚜蝇科Syrphidae、管蚜蝇亚科Eristalinae、管蚜蝇族Eristalini的一新属:艳管蚜蝇属Pseudomeromacrus gen.nov.和新种 刺茎艳管蚜蝇Pseudomeromacrus setipenitus Li,sp.nov.模式标本保存于华南农业大学植保系昆虫标本室。     

(蜓)类旋壁超微构造的研究——以麦(蜓)型旋壁为例

光学显微镜下的观察表明(蜒)类的旋壁属于钙质微粒壳类型,对它们的电镜观察揭示了组成旋壁的晶粒的大小、形态和排列方式.对具有致密层和蜂巢层型两层式旋壁的Triticites的电镜研究表明:其致密层由近等轴晶无定向排列而成;蜂巢层由短柱状晶定向排列组成板状复合体,进而组合成管状孔,镶嵌在不定向排列的近等轴品基质中构成.Pseudofusulina的致密层也由近等轴晶不定向排列构成,蜂巢层由粗柱晶定向排列组成纤状复合体,进而形成管状孔,镶嵌在近等轴晶基质中.