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61.
P69 is an isozyme of the medium size class of human 2'-5' oligoadenylate synthetases. In this study, recombinant P69 was expressed and used for enzymological and structural investigations. Bacterially expressed P69 was inactive whereas the same protein expressed in insect cells was highly active. Whether this difference could be due to differential post-translational modifications of the protein was investigated. Mutations of appropriate residues showed that myristoylation of the protein was not necessary for enzyme activity. In contrast, inhibition of glycosylation of P69, by tunicamycin treatment of the insect cells, produced an enzymatically inactive protein. Recombinant P69 produced in insect cells was purified by affinity chromatography. It was a dimeric glycoprotein, very stable and completely dependent on double stranded (ds) RNA for activity. The enzyme catalyzed the non-processive synthesis of 2'-5'-linked oligoadenylate products containing up to 30 residues. 2'-O-Methylated dsRNA was incapable of activating P69 and a 25-base pair dsRNA was as effective as larger dsRNA. This expression system will be useful for large scale production of P69 and its mutants for structural studies. 相似文献
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The lung is comprised of about 40 different cell types, of which only 15% are type II cells. These are the major, if not the sole, source of synthesis and secretion of lung surfactant. To date a large number of methods have been described for the isolation of pure populations of type II cells using a wide variety of techniques, but most of these have employed differential centrifugation methods and have used adult rodents. The present study reports the isolation of type II cells from fetal rabbit lungs by the immunoglobin G plating method. Pure populations of fetal type II cells in high yield and with good viability were obtained by the procedure for the first time from rabbit fetal tissue. 相似文献
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SN Popova MI Shadrina RI Fatkhlislamova EK Khusnutdinova PA Slominski? 《Canadian Metallurgical Quarterly》1998,34(6):843-845
A comparative study of two techniques for the PCR genotyping of highly polymorphic tandem repeats was carried out by the example of a triplet repeat in the myotonin protein kinase gene. Sequencing denaturing gels were shown to yield more precise results in the analysis of amplification products. 相似文献
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A Ziolkowski AS Shashkov AS Swierzko SN Senchenkova FV Toukach M Cedzynski KI Amano W Kaca YA Knirel 《Canadian Metallurgical Quarterly》1997,411(2-3):221-224
Structures of the O-specific polysaccharide chains of lipopolysaccharides from Proteus group OX strains (serogroups O1-O3) used as antigens in Weil-Felix test for diagnosis of rickettsiosis, were established. From them, the acid-labile polysaccharide of Proteus vulgaris OX19 (O1) is built up of the following branched pentasaccharide repeating units connected via a phosphate group: [structure in text] where QuiNAc stands for 2-acetamido-2,6-dideoxyglucose (N-acetylquinovosamine). The basis of serospecificity of the Proteus group OX antigens and their cross-reactivity with human anti-rickettsial antibodies is discussed. 相似文献
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Mating success of large and small flies of Drosophila malerkotliana and D. bipectinata was studied using multiple, male and female choice methods. In multiple choice method the large male mated with large female and small male paired with small female. In female choice method, large male was successful in mating with female irrespective of its size while in male choice method, preferential mating occurred between large male and large female as well as small male and small female. The consequence of such non-random mating in the presence of male rivarly and preferential mating has been discussed. 相似文献
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