RESUMEN
A novel actinomycete, designated strain KLBMP S0043(T), was isolated from the rhizosphere soil of Artemisia Linn. collected from the coastal region of Lianyungang, Jiangsu Province, in east China and was studied in detail for its taxonomic position. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain KLBMP S0043(T) is a member of the genus Nocardia. The 16S rRNA gene sequence similarity indicated that strain KLBMP S0043(T) is closely related to Nocardia asteroides NBRC 15531(T) (97.61 %) and Nocardia neocaledoniensis SBHR OA6(T) (97.38 %); similarity to other type strains of the genus Nocardia was found to be less than 97.2 %. The organism has chemical and morphological features consistent with its classification in the genus Nocardia such as meso-diaminopimelic acid as the diagnostic diamino acid in the cell wall peptidoglycan and arabinose and galactose as the diagnostic sugars. The predominant menaquinone was identified as MK-8(H4ω-cycl). Mycolic acids were detected. The diagnostic phospholipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides. The predominant cellular fatty acids were identified as C16:0, C18:0, C18:1ω9c, 10-methyl C18:0 [tuberculostearic acid (TBSA)] and summed feature 3 (C16:1ω7c/C16:1ω6c). The G+C content of the genomic DNA was determined to be 71.4 mol%. The results of DNA-DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of the strain from its most closely related strains. Based on morphological, chemotaxonomic and phylogenetic data, strain KLBMP S0043(T) is considered to represent a novel species of the genus Nocardia, for which the name Nocardia rhizosphaerae sp. nov. is proposed. The type strain is KLBMP S0043(T) (=CGMCC 4.7204 (T) = KCTC 29678(T)).
Asunto(s)
Nocardia/clasificación , Nocardia/aislamiento & purificación , Rizosfera , Microbiología del Suelo , Artemisia/crecimiento & desarrollo , Técnicas de Tipificación Bacteriana , Composición de Base , Carbohidratos/análisis , Pared Celular/química , China , Análisis por Conglomerados , Citosol/química , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácido Diaminopimélico/análisis , Ácidos Grasos/análisis , Microscopía Electrónica de Rastreo , Datos de Secuencia Molecular , Nocardia/genética , Nocardia/fisiología , Hibridación de Ácido Nucleico , Peptidoglicano/análisis , Fosfolípidos/análisis , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Vitamina K 2/análisisRESUMEN
AIM: To study the efficacy of heat shock protein 65 kDa (HSP65) of Mybobacterium tuberculosis var. bovis in prevention of autoimmune diabetes by intranasal. METHODS: The HSP65 gene was derived from Mybobacterium tuberculosis var. bovis genome by PCR and successfully expressed as soluble protein in Escherichia coli. The recombinant protein HSP65 was purified by anion exchange column chromatography, then used to immunize prediabetic NOD (non-obese diabetic) mice via three intranasal (i.n.) delivery in absence of adjuvants. Serum samples from the immunized mice were collected at monthly intervals. The anti-HSP65 antibody was detected by enzyme-linked immunosorbent assay (ELISA) and verified by Western blot analysis. The concentration of blood glucose was measured by automatic analyzer. RESULTS: Specific anti-HSP65 antibodies were successfully induced in mice immunized via intranasal routes. Histochemical analysis of mice pancreas tissue showed that HSP65 intranasal vaccination could decrease pathological changes in NOD mice. CONCLUSION: Intranasal vaccination with HSP65 in NOD mice could prevent the development of diabetes. Our results demonstrate that intranasal vaccination with HSP65 reduces significantly the inflammatory process associated with auto-immune diabetes. This approach may offer novel therapeutic avenues for the treatment for of type 1 diabetes mellitus.