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111.
The processing of macronuclear-destined DNA sequences microinjected into the macronuclear anlagen of the hypotrichous ciliate Stylonchia lemnae. 总被引:1,自引:1,他引:0
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We describe the construction of a vector carrying the micronuclear versions of two macronuclear DNA molecules, one of which was modified by the insertion of a polylinker sequence. This vector was injected into the polytene chromosomes of the developing macronucleus of Stylonychia and its processing during further macronuclear development and its fate in the mature macronucleus were analyzed. In up to 30% of injected cells the modified macronuclear DNA sequence could be detected. While the internal eliminated sequences (IES) present in the macronuclear precursor DNA sequence are still retained in the mature macronucleus, the modified macronuclear DNA sequence is correctly cut out from the vector, telomeres are added de novo and it is stably retained in the macronucleus during vegetative growth of the cells. This vector system represents an experimental system that allows the identification of DNA sequences involved in the processing of macronuclear DNA sequences during macronuclear development. 相似文献
112.
Wei Wen Su 《Biotechnology Techniques》1995,9(4):259-264
Summary The effect of agitation and aeration on filtration of Anchusa officinalis culture in a stirred tank bioreactor integrated with an internal filter unit was investigated. Increases in suction head of the pump that drove the filtration process were measured at impeller speeds of 100 and 200 rpm. Surprisingly, suction head attained at 200 rpm was about 40% higher than at 100 rpm. Direct observation of the cake deposition process in the reactor using a dilute cell suspension revealed that the filter cake formed at 100 rpm was thicker, but less compact. Aeration at 0.4 vvm was shown to have little effect on the filtration rate, since the bulk fluid flow was dominated by the impeller hydrodynamics. The initial flux can be recovered by filter backwashing with compressed air at a flow rate of 0.6 vvm for a duration of 5 minutes. 相似文献
113.
Summary Three screening methods were used to isolate GL-7-ACA acylase-producing strains. Three positive isolates were identified with Pseudomonas nitroreducens CCRC 11041 possessing the highest activity, against GL-7-ACA and GL-7-ADCA. No activity was detected when Ceph C or succinyl-7-ACA was used as substrate; glutaric acid was found to be inhibitory. CCRC 11041 could produce maximal GL-7-ACA acylase activity when cultivated on meat extract medium II. The enzyme had a pH optimum of 5.0 and a temperature optimum of 42°C. 相似文献
114.
Summary A simple and convenient technique was developed based on the principle of Warburg manometric method to measure O2 uptake rate (OUR) and CO2 evolution rate (CER) of suspended cells in a shake flask culture. It was successfully applied to suspension cultures of rice (Oryza sativa) and Panax notoginseng cells, and some important bioprocess parameters, such as OUR, CER, respiratory quotient (RQ), specific OUR (SOUR) and specific CER (SCER), were quantitatively obtained. The measuring system is easy to operate, able to treat many samples simultaneously and is economical. 相似文献
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117.
以克山病病区粮配成基础饲料,另在基础饲料中分别补充Se或VE,或Se+VE喂养大鼠,在细胞及亚细胞水平上以Ca代谢为主研究并比较了Se和VE在克山病病因中的作用。测量了心肌细胞和心肌线粒体的Ca代谢及有关指标、心肌线粒体能量转换功能及心肌组织自由基含量。结果表明,在低Se病区粮中补充Se或VE均能在一定程度上预防病区粮中致病因素对心肌细胞及线粒体的损伤;并且补充Se或VE均能使心肌组织中自由基含量减少。提示Se和VE是通过清除体内过量自由基预防细胞和线粒体的损伤的。但值得注意的是,实验中所用病区粮VE含量不低于甚至高于非病区对照粮,在低Se情况下,所补VE的量需要相当大(如本实验中补充200μg/g)才能较明显地预防心肌细胞和心肌线粒体的损伤。通过对这些结果的分析,进一步肯定低Se是克山病形成的重要因素之一。 相似文献
118.
Labeling of sialoglycoproteins with carbon-14 in vitro was performed by reacting the aldehyde groups, generated by mild periodate oxidation of the terminal sialyl groups, with 14C-labeled sodium cyanide to produce the labeled cyanohydrin derivatives (Kiliani reaction). Labeling with tritium was carried out by reduction of the aldehyde groups generated on the sialyl residues with 3H-labeled sodium borohydride following standard procedures. The behavior of both types of labeled specimens of fetuin and ovine submaxillary mucin, individually and in mixtures, was investigated by gel-filtration chromatography, gel electrophoresis, and cesium bromide gradient ultracentrifugation. The labeled sialyl residues were subjected to partial characterization: color yield with the resorcinol and thiobarbituric acid reagents, behavior on ion-exchange chromatography, and susceptibility to mild acid and enzymatic hydrolyses. In addition to these model glycoproteins, this procedure was also utilized to label the sialoglycoproteins present in human tracheobronchial secretions collected from normal subjects and patients with chronic bronchitis. The potential uses of this approach for comparative studies of normal and pathological sialoglycoconjugates available in minute amounts is described. The extension of this approach to the labeling of the galactosyl and N-acetylgalactosaminyl moieties of glycoconjugates following treatment with galactose oxidase is outlined. 相似文献
119.
Y Wen K S Famulski E Carafoli 《Biochemical and biophysical research communications》1984,122(1):237-243
The Ca2+-dependent K+ permeability of heart sarcolemma vesicles was measured by following the transmembrane movement of the charge compensating tetraphenylborate anion. The increase in vesicles permeability induced by Ca2+ is lost when membrane proteins are dephosphorylated by an endogenous protein phosphatase and is restored by a phosphorylation process catalysed by a cAMP-dependent protein kinase. The calmodulin antagonist R 24571 lowers the Ca2+-dependent K+ permeability by decreasing the Ca2+ affinity of the K+ transporting system. 相似文献
120.