Mutational effect of the “- 35” element of sorghumpsbA gene promoter

Mutations of the first position T and the third position G in TTGACA, the " - 35" element of sorghum psbA gene promoter, were induced using chemically synthesized 20 nt oligonucleotide primer. Three mutants were produced: ATTACA, GTGACA, and ATGACA. Then the protein binding affinity of the mutants and the wild type sorghum psbA gene promoter was tested in a spinach chloroplast protein extract system. Gel retardation assay of the wild type showed a strong protein-binding band. On the other hand, the protein-binding band of the mutant resulting from single base mutation, ATGACA or GTGACA, showed reduced intensity, while that of the mutant resulting from double base mutation, ATTACA, showed increased intensity. It is thus shown that the " - 35" element plays an important role in controlling the binding between psbA gene promoter and the specific chloroplast proteins; mutation of a single base may exert a substantial influence on the binding affinity.     

颗粒分布对金属基复合材料激光热冲击破坏的影响

从宏观和微观两方面研究了ＳｉＣ颗粒分布状况对陶瓷颗粒增强金属基复合材料激光热冲击破坏的影响,发现微裂纹形成是由基体内的孔洞和颗粒与基体脱胶所引起,而陶瓷颗粒分布不均匀对裂纹的扩展机理有重要影响,它大大降低了复合材料的宏观力学性能     

β葡聚糖酶杂合基因bglHAM的克隆及在P.pastoris中的表达

采用PCR的方法,以Bacillus macerans总DNA为模板,构建出β-1,3-1,4葡聚糖酶杂合基因bglHAM,与巴斯德毕赤酵母表达载体pPIC9K重组,得到重组质粒pPIC9K-HAM,电脉冲转化法转化酵母菌GS115,KM71,在MM,MD平板上筛选表型,YPD-G418平板上筛选多拷贝重组子,重组菌株经甲醇诱导后,刚果红平板染色法检测到β-葡聚糖酶活性,SDS-PAGE证明表达产物的分子量约为24kD,摇瓶诱导培养筛选到的重组菌株,胞外每mL发酵液最高酶活达240U。     

Transformation: a tool for studying fungal pathogens of plants

Plant diseases caused by plant pathogenic fungi continuously threaten the sustainability of global crop production. An effective way to study the disease-causing mechanisms of these organisms is to disrupt their genes, in both a targeted and random manner, so as to isolate mutants exhibiting altered virulence. Although a number of techniques have been employed for such an analysis, those based on transformation are by far the most commonly used. In filamentous fungi, the introduction of DNA by transformation typically results in either the heterologous (illegitimate) integration or the homologous integration of the transforming DNA into the target genome. Homologous integration permits a targeted gene disruption by replacing the wild-type allele on the genome with a mutant allele on transforming DNA. This process has been widely used to determine the role of newly isolated fungal genes in pathogenicity. The heterologous integration of transforming DNA causes a random process of gene disruption (insertional mutagenesis) and has led to the isolation of many fungal mutants defective in pathogenicity. A big advantage of insertional mutagenesis over the more traditional chemical or radiation mutagenesis procedures is that the mutated gene is tagged by transforming DNA and can subsequently be cloned using the transforming DNA. The application of various transformation-based techniques for fungal gene manipulation and how they have increased our understanding and appreciation of some of the most serious plant pathogenic fungi are discussed. Received 9 May 2001; received after revision 2 July 2001; accepted 3 July 2001     

Role of Cys221 and Asn116 in the zinc-binding sites of the Aeromonas hydrophila metallo-β-lactamase

The CphA metallo--lactamase produced by Aeromonas hydrophila exhibits two zinc-binding sites. Maximum activity is obtained upon binding of one zinc ion, whereas binding of the second zinc ion results in a drastic decrease in the hydrolytic activity. In this study, we analyzed the role of Asn116 and Cys221, two residues of the active site. These residues were replaced by site-directed mutagenesis and the different mutants were characterized. The C221S and C221A mutants were seriously impaired in their ability to bind the first, catalytic zinc ion and were nearly completely inactive, indicating a major role for Cys221 in the binding of the catalytic metal ion. By contrast, the binding of the second zinc ion was only slightly affected, at least for the C221S mutant. Mutation of Asn116 did not lead to a drastic decrease in the hydrolytic activity, indicating that this residue does not play a key role in the catalytic mechanism. However, the substitution of Asn116 by a Cys or His residue resulted in an approximately fivefold increase in the affinity for the second, inhibitory zinc ion. Together, these data suggested that the first zinc ion is located in the binding site involving the Cys221 and that the second zinc ion binds in the binding site involving Asn116 and, presumably, His118 and His196.Received 3 March 2003; received after revision 4 August 2003; accepted 25 August 2003     

紫外线诱变康宁木霉提高酶活力的研究

通过对康宁木霉菌种的诱变、筛选和固体发酵培养,研究了培养基、培养时间、诱变时间及pH值对诱变菌所产酶的活力的影响．结果表明,康宁木霉经诱变5min,在pH为6．0的玉米秸PDA培养基上培养72h,产生的C1酶和Cx酶的酶活力较高,C1酶的酶活力由73u／g提高到176u／g,增加了2．41倍,Cx酶的酶活力由798u／g提高到2069u／g,增加了2．59倍．     

换热器管束流体激振的研究现状评估及新方法

换热器管束的流体激振问题是严重影响其运行安全性的一个关键问题。该文对近年来国内外对此问题的研究进行了综述与评估。根据以往在叶轮机械流体激振研究方面开创的理论系统和研究方法,作者提出了通过应用振荡流体力学原理和参数多项式方法去求解流体基本方程,从而确定流体激振力的方法,应用全功能分析方法来准确预估流体弹性振动的稳定性边界,应用振荡压力传播理论来分析包括声共振在内的流体激振现象。这些思想为换热器管束流体激振研究指引了一条新途径。     

超冷玻色气体中的电磁感应透明

研究了超冷玻色气体中的电磁感应透明现象.利用平均场理论,考虑处于所有能级的原子被光势阱所囚禁,本文导出了系统的色散关系并计算了在超冷原子气体中传播的慢光的群速.     

3,4-Diphenylcycl[3,2,2]azine类化合物的合成及其光学性质研究

合成了3个不同取代基的3,4-Diphenylcycl[3,2,2]azine类化合物, 首次测试了它们各自的紫外光谱和荧光光谱,计算了它们的荧光量子效率和Stoke's位移,对比讨论了取代基效应对其紫外光谱和荧光光谱的影响,发现了其分子结构与光谱性质之间的某些规律性的关系.