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Optimized Transformation of Streptomyces sp. ATCC 39366 Producing Leptomycin by Electroporation
Yong-Qiang Fan,Hong-Jian Liu,Li Yan,Yu-Shi Luan,Hai-Meng Zhou,Jun-Mo Yang,Shang-Jun Yin,Yu-Long Wang 한국미생물학회 2013 The journal of microbiology Vol.51 No.3
Streptomyces sp. ATCC 39366 produces leptomycin derivatives. Leptomycin B, a potent and specific inhibitor against the export of nuclear proteins, is the main product; however, the introduction of DNA into this strain is almost impossible, which has impeded its further use. We developed a Streptomyces sp. ATCC 39366 transformation protocol to introduce foreign DNA via electroporation. Various conditions were examined, including treatments of the cell wall with weakening agents, electroporation parameters, and DNA content. We found that only plasmid DNA isolated from a dam- ET12567 strain resulted in successful transformation. The mycelium growing in a yeast-peptone-dextrose medium supplemented with 1% glycine at 28°C on a rotary shaker (220 rpm) was more dispersed than those without supplementation and prone to electroporation. The maximum transformation efficiency of 8×102 CFU/μg plasmid DNA was obtained at a field strength of 13 kV/cm with a time constant of 13 ms (25-μF capacitor; parallel resistance, 600 Ω) using 1-mm electrocuvettes. The results of the transformations of two other Streptomyces species indicated that the optimized conditions established in this study might only be applicable to Streptomyces sp. ATCC 39366. However, this is the first report of successful transformation of Streptomyces sp. ATCC 39366, and will facilitate the construction of a gene knockout mutant in Streptomyces sp. ATCC 39366 to produce series of new leptomycin derivatives.
Behavior of headed shear stud connectors subjected to cyclic loading
Fa-xing Ding,Guo-an Yin,Hai-bo Wang,Liping Wang,Qiang Guo 국제구조공학회 2017 Steel and Composite Structures, An International J Vol.25 No.6
The objective of this study is to investigate the actual behavior of studs in structures under earthquake load through laboratory tests and numerical simulation. A test program including eighteen specimens was devised with consideration of different concrete strengths and stud diameters. Six of specimens were subjected to monotonically increasing loading while the others were subjected to cyclic loading. Mechanical behavior including the failure mechanism, load-slip relationship, stiffness degradation, energy dissipation and the damage accumulation was obtained from the test results. An accurate numerical model based on the ABAQUS software was developed and validated against the test results. The results obtained from the finite element (FE) model matched well with the experimental results. Furthermore, based on the experimental and numerical data, the design formulas for expressing the skeleton curve were proposed and the simplified hysteretic model of load versus displacement was then established. It is demonstrated that the proposed formulas and simplified hysteretic model have a good match with the test results.
The diversity, recombination and horizontal transmission of Wolbachia in spiders in China
Yang Xiao-Hui,Gong Yu-Hui,Xu Xiang,Yin Hai-Qiang,Zhu Dao-Hong 한국응용곤충학회 2021 Journal of Asia-Pacific Entomology Vol.24 No.3
Wolbachia are maternally inherited endosymbiotic bacteria. These intracellular bacteria are common in ar thropods and could manipulate host reproduction in diverse ways, such as feminization, parthenogenesis, male killing and cytoplasmic incompatibility. In spiders, infection by Wolbachia has been found in a total of 99 species belonging to 62 genera and 17 families. Furthermore, recent studies analyzed the phylogeny of Wolbachia in Hylyphantes graminicola, 2 cave spiders and Agelenopsis species using multilocus sequence typing (MLST) approach. However, the diversity of Wolbachia strains determined by MLST in spiders from China is still largely unknown. In this study, we collected 1153 spider individuals from Mangshan in China and screened for Wolbachia in 975 individuals representing 68 spider species belonging to 45 genera of 16 families. We analyzed the phylogenetic relationship between Wolbachia and their host spiders by MLST approach. We found novel infections of Wol bachia in 1 family, 9 genera and 20 species of spiders. We found 13 new Wolbachia strains and suggest that group A is more common than group B in Wolbachia that infect spiders. Our results revealed three recombination events of the concatenated multilocus sequences in Wolbachia that infect spiders. Furthermore, our results demonstrated the phylogenetic incongruence between Wolbachia and spiders, suggesting the horizontal transmission of Wol bachia in spiders. We suggest that recombination and horizontal transmission may play an important role in the diversity and evolution of Wolbachia in spiders.