DNA sequence analysis of spontaneous histidine mutations in a polA1 strain of Escherichia coli K12 suggests a specific role of the GTGG sequence

Summary Spontaneously arising histidine mutations in an Escherichia coli K12 strain deficient for DNA polymerase I were analysed at the DNA sequence level. We screened approximately 150000 colonies and isolated 106 histidine auxotrophs. Of these, 98 were unstable hisC mutations; 12 representative mutants analysed were shown to have arisen by the excision of a single quadruplet repeat in the sequence 5INE" BORDER="0">-GCTGGCTGGCTGGCTG-3INE" BORDER="0">. Of the eight mutations at other sites, three hisA deletions and one hisD deletion occurred as a consequence of misalignment of tandemly repeated pentamers (hisD) or decamers (hisA). A single hisA point mutation was found to be a missense mutation. Two extended deletions, covering the his operon were not analysed. We could not identify the hisC deletion by sequencing. We conclude that polA1 is a strong imitator that induces mutations mostly of the minus frameshift and deletion type by a Streisinger-type of mispairing in repetitive DNA sequences. Finally, the possible role of a 5INE" BORDER="0">-GTGG-3INE" BORDER="0"> sequence and its inverted or direct complements, which are found in the vicinity of all the deletions and frameshifts, is discussed.     

绝经后女性血清瘦素、骨密度及血骨特异性碱性磷酸酶和Ⅰ型胶原交联氨基末端肽的测定及意义

目的:测定绝经后女性血清瘦素(leptin)与骨密度及及血清骨特异性碱性磷酸酶(BAP)和Ⅰ型胶原交联氨基末端肽(NTx)并探讨其关系。方法:用酶联免疫吸附试验测定287名40-80岁健康绝经后女性血清leptin以及血清骨特异性碱性磷酸酶(BAP)和Ⅰ型胶原交联氨基末端肽(NTx);用双能X线骨密度扫描仪测定总体、腰椎正位、总髋部骨密度以及体脂、瘦体重;分析它们之间的关系。结果:Leptin与髋部总体BMD呈正相关(r=0.162,P<0.05),校正年龄和体脂后,Leptin与髋部总体BMD相关性消失,Leptin与BAP相关无统计学意义;与NTX呈负相关(r=-0.119,P<0.05),校正年龄和体脂后,相关无统计学意义。BAP与总体骨密度、腰椎骨密度、髋部总体骨密度均呈负相关(r=-0.210,r=-0.236,r=-0.223,P<0.05),校正年龄和体质指数后,相关性都依然存在(r=-0.168,r=-0.187,r=-0.169,P<0.05)。NTx与总体骨密度、腰椎骨密度、髋部总体骨密度均呈负相关(r=-0.238,r=-0.232,r=-0.239,P<0.05),校正年龄和体质指数后...     

Hemocyanin gene family evolution in spiders (Araneae), with implications for phylogenetic relationships and divergence times in the infraorder Mygalomorphae

Hemocyanins are multimeric copper-containing hemolymph proteins involved in oxygen binding and transport in all major arthropod lineages. Most arachnids have seven primary subunits (encoded by paralogous genes a–g), which combine to form a 24-mer (4 × 6) quaternary structure. Within some spider lineages, however, hemocyanin evolution has been a dynamic process with extensive paralog duplication and loss. We have obtained hemocyanin gene sequences from numerous representatives of the spider infraorders Mygalomorphae and Araneomorphae in order to infer the evolution of the hemocyanin gene family and estimate spider relationships using these conserved loci. Our hemocyanin gene tree is largely consistent with the previous hypotheses of paralog relationships based on immunological studies, but reveals some discrepancies in which paralog types have been lost or duplicated in specific spider lineages. Analyses of concatenated hemocyanin sequences resolved deep nodes in the spider phylogeny and recovered a number of clades that are supported by other molecular studies, particularly for mygalomorph taxa. The concatenated data set is also used to estimate dates of higher-level spider divergences and suggests that the diversification of extant mygalomorphs preceded that of extant araneomorphs. Spiders are diverse in behavior and respiratory morphology, and our results are beneficial for comparative analyses of spider respiration. Lastly, the conserved hemocyanin sequences allow for the inference of spider relationships and ancient divergence dates.     

(+)-Nootkatone inhibits tumor necrosis factor α/interferon γ-induced production of chemokines in HaCaT cells

Chemokines are important mediators of cell migration, and thymus and activation-regulated chemokine (TARC/CCL17) and macrophage-derived chemokine (MDC/CCL22) are well-known typical inflammatory chemokines involved in atopic dermatitis (AD). (+)-Nootkatone is the major component of Cyperus rotundus. (+)-Nootkatone has antiallergic, anti-inflammatory, and antiplatelet activities. The purpose of this study was to investigate the effect of (+)-nootkatone on tumor necrosis factor α (TNF-α)/interferon γ (IFN-γ)-induced expression of Th2 chemokines in HaCaT cells. We found that (+)-nootkatone inhibited the TNF-α/IFN-γ-induced expression of TARC/CCL17 and MDC/CCL22 mRNA in HaCaT cells. It also significantly inhibited TNF-α/IFN-γ-induced activation of nuclear factor kappa B (NF-κB), p38 mitogen-activated protein kinase (MAPK), and protein kinase Cζ (PKCζ). Furthermore, we showed that PKCζ and p38 MAPK contributed to the inhibition of TNF-α/IFN-γ-induced TARC/CCL17 and MDC/CCL22 expression by blocking IκBα degradation in HaCaT cells. Taken together, these results suggest that (+)-nootkatone may suppress TNF-α/IFN-γ-induced TARC/CCL17 and MDC/CCL22 expression in HaCaT cells by inhibiting of PKCζ and p38 MAPK signaling pathways that lead to activation of NF-κB. We propose that (+)-nootkatone may be a useful therapeutic candidate for inflammatory skin diseases such as AD.     

Studying dynamic process of photosensitization by ESR method

By kinetic methods, functional relation of TAN radical, produced in the process of TEMPONE trapping O2, vs. time during photosensitization was established. Accordingly relative rate constants of generating all kinds of active intermediates through types I and II mechanism of photosensitization can be calculated. Using the formula and experimental results, the relative rate constants of generating O2, O2 and PS2 of three kinds of perylenequinone photosensitizer: HA, HB and CP in DMF-H20 and DMSO-H2O system were calculated, and then the structure-activity relationship of perylenequinone photosensitizer and the relation between photosensitivity and solvent was studied.     

Identification and functional characterization of phenylglycine biosynthetic genes involved in pristinamycin biosynthesis in Streptomyces pristinaespiralis

Pristinamycin I (PI), a streptogramin type B antibiotic produced by Streptomyces pristinaespiralis, contains the aproteinogenic amino acid l-phenylglycine. Recent sequence analysis led to the identification of a set of putative phenylglycine biosynthetic genes. Successive inactivation of the individual genes resulted in a loss of PI production. Production was restored by supplementation with externally added l-phenylglycine, which demonstrates that these genes are involved in phenylglycine biosynthesis and thus probably disclosing the last essential pristinamycin biosynthetic genes. Finally, a putative pathway for phenylglycine synthesis is proposed.     

Somatic hybridization between male sterile Nicotiana tabacum and N. glutinosa through protoplast fusion

Summary Protoplasts derived from suspension cultured cells of cytoplasmic male sterile Nicotiana tabacum (N. debneyi cytoplasm) and of fertile N. glutinosa were fused with the aid of polyethylene glycol (PEG). Out of 1,089 colonies developed from PEG-treated protoplasts, 29 restored whole plants.A somatic hybrid plant was selected on the basis of isoelectrofocusing analysis of Fraction I protein in leaves of regenerated plants. A newly created hybrid contained small subunits of both parents but only a N. glutinosa type large subunit.Male sterile character was conserved in a hybrid plant while leaf morphology was intermediate between the parents. By tobacco mosaic virus infection tests, the hybrid's leaves showed resistant symptoms, hypersensitive local lesions, which were due to N. glutinosa nuclear genome expression.Abbreviations PEG Polyethylene glycol - TMV Tobacco mosaic virus     

PCR防污染技术的研究进展

聚合酶链式反应（PCR）是一种高灵敏核酸扩增技术,广泛应用于核酸检测中。但在实际应用过程中,扩增产物及其他核酸片段的污染会导致假阳性的结果,制约了PCR在临床检测中的应用。为了解决这一问题,建立了许多PCR防污染的方法,除了早期建立的并已得到广泛应用的物理隔绝法、光照法及水解法外,近年来还发展了酶消化法、化学修饰法及DEAE纤维素法。本文对PCR防污染技术的原理、应用及进展进行了综述。     

主要组织相容性复合体I类分子抗原肽

被主要组织相容性复合体(MHC)I类分子呈递在细胞表面的抗原肽大部分来源于细胞内新合成蛋白质的降解产物,抗原肽直接体现细胞内功能蛋白质的部分变化,蛋白酶体、氨肽酶和抗原转运体(TAP)参与调控抗原肽的生成。在MHC的组装、折叠过程中,抗原肽促进各亚基的结合和折叠进程;而在起始细胞的免疫应答过程中,抗原肽不仅诱导T细胞抗原受体的特异结合,更为重要的是延长MHC同T细胞抗原受体特异结合的作用时间。     

P430: a retrospective, 1971–2001

The spectral species P430 and its spectral and kinetic properties are briefly reviewed. Currently available evidence shows P430 to be the optic-spectral representation of FeS-A/B, the electron acceptor(s) of Photosystem I (PS I). Retired (formerly with the Charles F. Kettering Research Laboratory, Yellow Springs, Ohio, USA). This revised version was published online in June 2006 with corrections to the Cover Date.