RESUMO
A recombinant scFv antibody against human spleen ferritin was expressed as a barstar-fused protein in Escherichia coli and in Nicotiana tabacum plants and suspension cell cultures. As demonstrated by immunoblotting with antibarstar antibodies, direction of the recombinant protein to the endomembrane system of plant cells ensured its stability and solubility. Production of the recombinant protein did not differ between parental transgenic plants and their first-generation progeny. Fusion with barstar allowed not only immunochemical detection of the recombinant scFv antibody, but also their purification from the plant material by affinity chromatography with barnase-His6 immobilized on a metal-affinity carrier.
Assuntos
Bactérias/imunologia , Proteínas de Bactérias , Ferritinas/imunologia , Fragmentos de Imunoglobulinas/biossíntese , Plantas/imunologia , Sequência de Bases , Cromatografia de Afinidade , Primers do DNA , Humanos , Proteínas/química , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/química , Ribonucleases/químicaRESUMO
Tobacco plants were genetically transformed with the Arabidopsis thaliana heterologous hmg1 gene encoding 3-hydroxy-3-methylglutaryl-CoA reductase, a key enzyme involved in the metabolism of terpenoid compounds. The hmg1 gene was inserted under the control of the 35S RNA double promoter from the cauliflower mosaic virus (CaMV 35S) both in direct and reverse orientation relative to the promoter. DNA analysis by polymerase chain reaction (PCR) and Southern blotting confirmed the transgenic nature of the tobacco plants obtained. DNA-RNA hybridization revealed expression of the hmg1 gene in these tobacco plants. The plants transformed with the antisense copy of the hmg1 gene differed from the control plants in delayed development and in flower color and shape.
Assuntos
Elementos Antissenso (Genética) , Hidroximetilglutaril-CoA Redutases/genética , Nicotiana/genética , Plantas Geneticamente Modificadas/genética , Plantas Tóxicas , Arabidopsis/genética , Sequência de Bases , Southern Blotting , Caulimovirus/genética , Primers do DNA , Regulação da Expressão Gênica de Plantas , Hidroximetilglutaril-CoA-Redutases NADP-Dependentes , Hibridização de Ácido Nucleico , Fenótipo , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas , Transformação GenéticaRESUMO
A comparative analysis of zono - and tomography in 69 patients with minor peripheral cancer of the lung (26), tuberculoma (24), hamartoma (19) was performed. Zonography was shown to improve differential diagnosis between them. However, its importance is limited and helps to better visualize changes of the lung tissue around the formations, "a path" outgoing to the pulmonary radix and chest wall, and to detect the enlarged intrathoracic lymph nodes. Tomography is of greater informative value for the evaluation of the skialogical features of the formations and pleural changes. The combined use of zono - and tomography results in a better detection of different pathological lung formations of small sizes and changes in the pulmonary tissue enhancing the efficacy of differential radiodiagnosis between them.