目的 建立逼尿肌细胞的原代培养及鉴定方法.方法 应用Ⅱ型胶原酶消化分离逼尿肌细胞,在含20%胎牛血清DMEM培养液中培养,观察细胞形态和扩增情况,用a-SM-Actin进行免疫组化鉴定细胞类型.结果 逼尿肌细胞在培养18 h 后开始贴壁在瓶底,4～5...目的 建立逼尿肌细胞的原代培养及鉴定方法.方法 应用Ⅱ型胶原酶消化分离逼尿肌细胞,在含20%胎牛血清DMEM培养液中培养,观察细胞形态和扩增情况,用a-SM-Actin进行免疫组化鉴定细胞类型.结果 逼尿肌细胞在培养18 h 后开始贴壁在瓶底,4～5 d 后可见细胞融合,8～10 d 后可见细胞覆盖瓶底80%以上.a-SM-Actin免疫组化染色鉴定,光镜下观察梭形细胞胞浆内见纵行排列的棕黄色丝状物.结论 该方法简单、易于掌握,短期内可获得大量高纯度的逼尿肌细胞.展开更多
It is a practical approach to select candidate probiotic bacterial stains on the basis of their special traits. Production of digestive enzyme was used as a trait to select a candidate probiotic bacterial strain in th...It is a practical approach to select candidate probiotic bacterial stains on the basis of their special traits. Production of digestive enzyme was used as a trait to select a candidate probiotic bacterial strain in this study. In order to select a bacterium with the ability to degrade both starch and protein, an ideal bacterial strain STE was isolated from marine shrimp (Litopenaeus vannamei) intestines by using multiple selective media.The selected isolate STE was identified on the basis of its morphological, physiological,and biochemical characteristics as well as molecular analyses. Results of degradation experiments confirmed the ability of the selected isolate to degrade both starch and casein. The isolate STE was aerobic, Gram-negative, rod-shaped, motile and non-spore-forming, and had catalase and oxidase activities but no glucose fermentation activity. Among the tested carbon/nitrogen sources, only Tween40, alanyl-glycine, aspartyl-glycine, and glycyl-l-glutamic acid were utilized by the isolate STE. Results of homology comparison analyses of the 16S rDNA sequences showed that the isolate STE had a high similarity to several Pseudoalteromonas species and, in the phylogenetic tree, grouped with P. ruthenica with maximum bootstrap support (100%). In conclusion, the isolate STE was characterized as a novel strain belonging to the genus Pseudoalteromonas.This study provides a further example of a probiotic bacterial strain with specific characteristics isolated from the host gastrointestinal tract.展开更多
A crude protease produced from Planomicrobium sp. L-2 is described, and its effectiveness as an additive in liquid detergent evaluated. We isolate the protease-producing Planomicrobium sp. L-2 from the gastrointestina...A crude protease produced from Planomicrobium sp. L-2 is described, and its effectiveness as an additive in liquid detergent evaluated. We isolate the protease-producing Planomicrobium sp. L-2 from the gastrointestinal tract of Octopus variabilis. At least three caseinolytic protease clear bands were observed in zymogram analysis. The crude alkaline protease was highly tolerant of a pH range from 7.0 to 9.0, and temperatures to 50℃ after incubation for 1 h. Proteolytic enzymes were stable towards three surfactants （5% Tween 80, 1% Triton X-100 and 0.05% SDS） and an oxidizing agent （1% hydrogen peroxide）, in addition to being highly stable and compatible with popular commercial laundry powered detergent brands available in China. Our study demonstrates the potential these proteases have for development into novel classes of detergent additive. This study also suggests that the gastrointestinal tract of Octopus variabilis may be a rich source of commercially valuable strains of enzvme.展开更多
文摘目的 建立逼尿肌细胞的原代培养及鉴定方法.方法 应用Ⅱ型胶原酶消化分离逼尿肌细胞,在含20%胎牛血清DMEM培养液中培养,观察细胞形态和扩增情况,用a-SM-Actin进行免疫组化鉴定细胞类型.结果 逼尿肌细胞在培养18 h 后开始贴壁在瓶底,4～5 d 后可见细胞融合,8～10 d 后可见细胞覆盖瓶底80%以上.a-SM-Actin免疫组化染色鉴定,光镜下观察梭形细胞胞浆内见纵行排列的棕黄色丝状物.结论 该方法简单、易于掌握,短期内可获得大量高纯度的逼尿肌细胞.
基金financially supported by the National High Technology Research and Development Project of China(863 Project,No.2001AA622060 and No.2003A-A622060)
文摘It is a practical approach to select candidate probiotic bacterial stains on the basis of their special traits. Production of digestive enzyme was used as a trait to select a candidate probiotic bacterial strain in this study. In order to select a bacterium with the ability to degrade both starch and protein, an ideal bacterial strain STE was isolated from marine shrimp (Litopenaeus vannamei) intestines by using multiple selective media.The selected isolate STE was identified on the basis of its morphological, physiological,and biochemical characteristics as well as molecular analyses. Results of degradation experiments confirmed the ability of the selected isolate to degrade both starch and casein. The isolate STE was aerobic, Gram-negative, rod-shaped, motile and non-spore-forming, and had catalase and oxidase activities but no glucose fermentation activity. Among the tested carbon/nitrogen sources, only Tween40, alanyl-glycine, aspartyl-glycine, and glycyl-l-glutamic acid were utilized by the isolate STE. Results of homology comparison analyses of the 16S rDNA sequences showed that the isolate STE had a high similarity to several Pseudoalteromonas species and, in the phylogenetic tree, grouped with P. ruthenica with maximum bootstrap support (100%). In conclusion, the isolate STE was characterized as a novel strain belonging to the genus Pseudoalteromonas.This study provides a further example of a probiotic bacterial strain with specific characteristics isolated from the host gastrointestinal tract.
基金Supported by the Natural Science Foundation of Shandong Province(No.ZR2011CM023)the High-Level Personnel Research Foundation of Qingdao Agricultural University(No.631431)
文摘A crude protease produced from Planomicrobium sp. L-2 is described, and its effectiveness as an additive in liquid detergent evaluated. We isolate the protease-producing Planomicrobium sp. L-2 from the gastrointestinal tract of Octopus variabilis. At least three caseinolytic protease clear bands were observed in zymogram analysis. The crude alkaline protease was highly tolerant of a pH range from 7.0 to 9.0, and temperatures to 50℃ after incubation for 1 h. Proteolytic enzymes were stable towards three surfactants （5% Tween 80, 1% Triton X-100 and 0.05% SDS） and an oxidizing agent （1% hydrogen peroxide）, in addition to being highly stable and compatible with popular commercial laundry powered detergent brands available in China. Our study demonstrates the potential these proteases have for development into novel classes of detergent additive. This study also suggests that the gastrointestinal tract of Octopus variabilis may be a rich source of commercially valuable strains of enzvme.
