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61.
Drum CL Yan SZ Sarac R Mabuchi Y Beckingham K Bohm A Grabarek Z Tang WJ 《The Journal of biological chemistry》2000,275(46):36334-36340
The edema factor exotoxin produced by Bacillus anthracis is an adenylyl cyclase that is activated by calmodulin (CaM) at resting state calcium concentrations in infected cells. A C-terminal 60-kDa fragment corresponding to the catalytic domain of edema factor (EF3) was cloned, overexpressed in Escherichia coli, and purified. The N-terminal 43-kDa domain (EF3-N) of EF3, the sole domain of edema factor homologous to adenylyl cyclases from Bordetella pertussis and Pseudomonas aeruginosa, is highly resistant to protease digestion. The C-terminal 160-amino acid domain (EF3-C) of EF3 is sensitive to proteolysis in the absence of CaM. The addition of CaM protects EF3-C from being digested by proteases. EF3-N and EF3-C were expressed separately, and both fragments were required to reconstitute full CaM-sensitive enzyme activity. Fluorescence resonance energy transfer experiments using a double-labeled CaM molecule were performed and indicated that CaM adopts an extended conformation upon binding to EF3. This contrasts sharply with the compact conformation adopted by CaM upon binding myosin light chain kinase and CaM-dependent protein kinase type II. Mutations in each of the four calcium binding sites of CaM were examined for their effect on EF3 activation. Sites 3 and 4 were found critical for the activation, and neither the N- nor the C-terminal domain of CaM alone was capable of activating EF3. A genetic screen probing loss-of-function mutations of EF3 and site-directed mutations based on the homology of the edema factor family revealed a conserved pair of aspartate residues and an arginine that are important for catalysis. Similar residues are essential for di-metal-mediated catalysis in mammalian adenylyl cyclases and a family of DNA polymerases and nucleotidyltransferases. This suggests that edema factor may utilize a similar catalytic mechanism. 相似文献
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五唇兰雌配子体发育和胚胎发生的研究 总被引:11,自引:2,他引:11
五唇兰的胚珠倒生型,具薄珠心,两层珠被。胚囊发育为双孢子葱型,成熟胚囊8核。从传粉到受精约50d,正常双受精。胚具5-6细胞的胚柄,种子成熟时胚柄及胚乳核消失,成熟种子只具单层细胞的种皮和一个未分化的珠珠形胚。 相似文献
65.
Yan-Ru Cao Yi Jiang Qian Wang Shu-Kun Tang Wen-Xiang He Quan-Hong Xue Li-Hua Xu Cheng-Lin Jiang 《Antonie van Leeuwenhoek》2010,98(3):389-394
A novel pink-coloured, non-spore-forming, non-motile, Gram-negative bacterium, designated YIM 48858T, is described by using a polyphasic approach. The strain can grow at pH 6.5–9 (optimum at pH 7) and 25–30°C (optimum at 28°C).
NaCl is not required for its growth. Positive for oxidase and catalase. Urease activity, nitrate reduction, starch and Tween
80 tests are negative reaction. 16S rRNA gene sequence similarity studies showed that strain YIM 48858T is a member of the genus Rubellimicrobium, with similarities of 96.3, 95.7 and 95.5% to Rubellimicrobium mesophilum MSL-20T, Rubellimicrobium aerolatum 5715S-9T and Rubellimicrobium
thermophilum DSM 16684T, respectively. Q-10 was the predominant respiratory ubiquinone as in the other members of the genus Rubellimicrobium. The major polar lipids were diphosphatidylglycerol, phosphatidylcholine, phosphoglycolipid, glycolipid and the major fatty
acids were C18:1 ω7c, C16:0 and C10:0 3-OH, which are very different from the valid published species. The DNA G + C content
was 67.7 mol%. Both phylogenetic and chemotaxonomic evidence supports that YIM 48858T is a novel species of the genus Rubellimicrobium, for which the name Rubellimicrobium roseum sp. nov. is proposed. The type strain is YIM 48858T (=CCTCC AA 208029T =KCTC 23202T). 相似文献
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Xian Qi Yuning Pan Yuanfang Qin Rongqiang Zu Fengyang Tang Minghao Zhou Hua Wang Yongchun Song 《中国病毒学》2012,27(5):292-298
Currently, three predominant subtypes of influenza virus are prevalent in pig populations worldwide: H1N1, H3N2, and H1N2. European avian-like H1N1 viruses, which were initially detected in European pig populations in 1979, have been circulating in pigs in eastern China since 2007. In this study, six influenza A viruses were isolated from 60 swine lung samples collected from January to April 2011 in eastern China. Based on whole genome sequencing, molecular characteristics of two isolates were determined. Phylogenetic analysis showed the eight genes of the two isolates were closely related to those of the avian-like H1N1 viruses circulating in pig populations, especially similar to those found in China. Four potential glycosylation sites were observed at positions 13, 26, 198, 277 in the HA1 proteins of the two isolates. Due to the presence of a stop codon at codon 12, the isolates contained truncated PB1-F2 proteins. In this study, the isolates contained 591Q, 627E and 701N in the polymerase subunit PB2, which had been shown to be determinants of virulence and host adaptation. The isolates also had a D rather than E at position 92 of the NS1, a marker of mammalian adaptation. Both isolates contained the GPKV motif at the PDZ ligand domain of the 3′ end of the NS1, a characteristic marker of the European avian-like swine viruses since about 1999, which is distinct from those of avian, human and classical swine viruses. The M2 proteins of the isolates have the mutation (S31N), a characteristic marker of the European avian-like swine viruses since about 1987, which may confer resistance to amantadine and rimantadine antivirals. Our findings further emphasize the importance of surveillance on the genetic diversity of influenza A viruses in pigs, and raise more concerns about the occurrence of cross-species transmission events. 相似文献
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丝状真菌(Filamentous fungi)的发酵生产通常具有较高的工业应用价值,但其菌体形态是一个区别于其他非丝状菌的一个重要发酵指标。针对目前形态分析的瓶颈,本研究使用琼脂糖凝胶对黑曲霉菌形进行固定,利用平板实现菌球样本的大量制备,并结合图形处理软件自建自动化处理程序,实现了大量准确可靠的菌体形态参数的获得,大大增加了形态数据处理通量及准确度。应用该方法于黑曲霉发酵生产糖化酶过程中不同供氧水平及剪切水平下菌体形态的研究,通过大量形态数据定量阐明了黑曲霉在不同剪切水平下的分区域形态分布特性,为进一步工业过程的形态优化提供了重要的研究方法。 相似文献