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41.
A simple and efficient procedure for the isolation of high-quality phage lambda DNA using a DEAE-cellulose column 总被引:4,自引:0,他引:4
A simple and rapid procedure for purifying large quantities of bacteriophage lambda particles and DNA is described. The procedure involves DEAE-cellulose column chromatography of the phage particles and elution of the phage particles from the column with a low-ionic-strength buffer. The resulting phage were well separated from RNA, DNA, and proteins derived from Escherichia coli host cells. The lambda DNA was prepared from the purified phage particles by the conventional method of phenol extraction and ethanol precipitation. This procedure did not use nucleases, proteases, detergents, or CsCl density gradient centrifugation. The lambda DNA obtained by this method was equivalent in purity to the material prepared by CsCl density gradient centrifugation and amenable to restriction enzyme digestion, ligation, radiolabeling, and double-stranded DNA sequencing. A detailed protocol is described for obtaining 0.5 to 1.0 mg DNA from a 1-liter liquid lysate in less than 5 h. This procedure is simple, inexpensive, and timesaving, and is particularly suitable for large-scale isolation of lambda DNA. 相似文献
42.
Tomoo Suzuki Yoshihiro Ichihara Masaru Yamada Kenzo Tonomura 《Bioscience, biotechnology, and biochemistry》2013,77(4):747-756
Pseudomonas 0–3 strain which was isolated from soil can grow on polyvinyl alcohol (PVA) as a sole carbon source. When 0.5 per cent of PVA (500, 1500 or 2000) was employed as the carbon source in the culture medium, PVA was almost completely lost from the culture fluid after a week and the concentration of total organic carbon measured by a TOC analyzer decreased from the initial value of about 2700 ppm to 250~300 ppm after 7~10 days culture. This bacterium was found to produce and secrete an inducible enzyme which degrade PVA. The way by which this enzyme degrades PVA was examined and the results were obtained which suggested that PVA was broken down oxidatively in a way of endowise splitting. However, the mechanism of PVA degradation has not been clarified yet. The optimum pH and temperature for enzyme activity were examined and they were 7.5~8.5 and 35~45°C, respectively. Morphological and biological characteristics of this bacterium were examined and it was similar to a strain of Pseudomonas boreopolis. 相似文献
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44.
Taichiro Miyake Kosuke Soda Yasushi Itoh Yoshihiro Sakoda Hirohito Ishigaki Tomoya Nagata Hideaki Ishida Misako Nakayama Hiroichi Ozaki Hideaki Tsuchiya Ryuzo Torii Hiroshi Kida & Kazumasa Ogasawara 《Journal of medical primatology》2010,39(1):58-70
Background Highly pathogenic avian influenza virus (HPAIV) infection has a high mortality rate in humans. Secondary bacterial pneumonia with HPAIV infection has not been reported in human patients, whereas seasonal influenza viruses sometimes enhance bacterial pneumonia, resulting in substantial morbidity and mortality. Therefore, if HPAIV infection were accompanied by bacterial infection, an increase in mortality would be expected. We examined whether a vaccine against HPAIV prevents severe morbidity caused by mixed infection with HPAIV and bacteria.
Methods H7N7 subtype of HPAIV and Streptococcus pneumoniae were inoculated into cynomolgus macaques with or without vaccination of inactivated whole virus particles .
Results Vaccination against H7N7 HPAIV decreased morbidity caused by HPAIV and pneumonia caused by S. pneumoniae . Bacterial replication in lungs was decreased by vaccination against HPAIV, although the reduction in bacterial colonies was not significant.
Conclusions Vaccination against HPAIV reduces pneumonia caused by bacterial superinfection and may improve prognosis of HPAIV-infected patients. 相似文献
Methods H7N7 subtype of HPAIV and Streptococcus pneumoniae were inoculated into cynomolgus macaques with or without vaccination of inactivated whole virus particles .
Results Vaccination against H7N7 HPAIV decreased morbidity caused by HPAIV and pneumonia caused by S. pneumoniae . Bacterial replication in lungs was decreased by vaccination against HPAIV, although the reduction in bacterial colonies was not significant.
Conclusions Vaccination against HPAIV reduces pneumonia caused by bacterial superinfection and may improve prognosis of HPAIV-infected patients. 相似文献
45.
Hajime Sakaguchi Hiromi Hagiwara Yoshihiro Fukumori Yutaka Tamaura Minoru Funaki Shigehisa Hirose 《FEMS microbiology letters》1993,107(2-3):169-173
Abstract The magnetic bacterium Magnetospirillum magnetotacticum prefers a microaerobic habitat and should be able to sense oxygen. Therefore, the bacterium was cultured under atmospheres containing 0–5% O2 and analyzed for oxygen-dependent changes in the levels of its protein components by sodium dodecyl sulfate-polyccrylamide gel electrophoresis (SDS-PAGE). The analysis revealed a marked anaerobic induction of a 140-kDa protein, which was suppressed when M. magnetotacticum was switched from microaerobic (<1% O2 ) to aerobic (>1% O2 ) growth conditions. Although its function remains to be determined, the 140-kDa protein may serve as a useful tool to gain insight into the physiology of the organism. 相似文献
46.
To study the participation of chloroplast protein synthesisduring the three phases [Matsuda (1974) Biochim. Biophys. Acta366:45] of the greening process in Chlamydomonas reinhardtiiy-1, the greening characteristics in the low-chloroplast ribosomemutant y-1 ac-20 were compared with those in the y-1. In thedouble mutant cells Chl synthesis proceeded with an extendedlag and without a second transition point. The development ofpotential for rapid Chl synthesis (P-factor formation) was alsodelayed. Furthermore, PS I activity increased significantly,whereas PS II activity developed very little during greeningof the double mutant cells. The results indicate that greeningin double mutant cells occurs with no apparent late phase. (Received November 26, 1984; Accepted February 25, 1985) 相似文献
47.
Mori Hideki Masaki Takashi Tsunamoto Yoshihiro Naoe Shoji 《Journal of plant research》2020,133(2):193-203
Journal of Plant Research - Lianas have a huge influence on forest structure and function. However, it is unclear how the surrounding environment affects the establishment of liana seedlings in... 相似文献
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49.
Fumiya Noguchi Masaru Kawato Takao Yoshida Yoshihiro Fujiwara Katsunori Fujikura Kiyotaka Takishita 《The Journal of eukaryotic microbiology》2013,60(2):158-165
It has recently been unveiled that a wide variety of microbial eukaryotes (protists) occur in chemosynthetic ecosystems, such as hydrothermal vents and methane seeps. However, there is little knowledge regarding protists associated with endemic animals inhabiting these environments. In the present study, utilizing PCR techniques, we detected fragments of the small subunit ribosomal RNA gene (SSU rRNA gene) from a particular protist from gill tissues of a significant fraction of the vesicomyid clams Calyptogena soyoae and C. okutanii complex and of the mussel Bathymodiolus platifrons and B. japonicus, all of which harbor chemosynthetic endosymbiont bacteria and dominate methane seeps in Sagami Bay, Japan. Based on the phylogeny of SSU rRNA gene, the organism in question was shown to belong to Alveolata. It is noteworthy that this protist did not affiliate with any known alveolate group, although being deeply branched within the lineage of Syndiniales, for which the monophyly was constantly recovered, but not robustly supported. In addition, the protist detected using PCR followed by sequencing was localized within gill epithelial cells of B. platifrons with whole‐mount fluorescence in situ hybridization. This protist may be an endoparasite or an endocommensal of Calyptogena spp. and Bathymodiolus spp., and possibly have physiological and ecological impacts on these bivalves. 相似文献
50.
Kenichi Tanaka Masaaki Nakayama Makoto Kanno Hiroshi Kimura Kimio Watanabe Yoshihiro Tani Yuki Kusano Hodaka Suzuki Yoshimitsu Hayashi Koichi Asahi Keiji Sato Toshio Miyata Tsuyoshi Watanabe 《PloS one》2013,8(12)