RESUMO
A novel Gram-staining-negative bacterium, designated DH-5T, was isolated from a farmland soil in Chuzhou, Anhui province, China. Cells of strain DH-5T were aerobic, non-motile, non-spore-forming and rod-shaped. The organism grew at 20-37 °C, pH 6.0-9.0 and with 0-5 % NaCl (w/v). The DNA G+C content was 42.8 mol%. The major fatty acids (>5 %) were iso-C15 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), iso-C17 : 0 3-OH and C16 : 0. The respiratory quinone was MK-7, and the major polar lipids were phosphatidylethanolamine and phosphoglycolipid. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain DH-5T was a member of the genus Sphingobacterium and shared the highest similarity with Sphingobacterium gobiense H7T (96.0 %), followed by Sphingobacterium arenae H-12T (94.5 %). Strain DH-5T exhibited low DNA-DNA relatedness with S. gobiense H7T (35.1±1.4 %) and S. arenae H-12T (21.4±1.0 %). On the basis of phenotypic, genotypic and phylogenetic evidence, DH-5T is considered to represent a novel species of the genus Sphingobacterium, for which the name Sphingobacterium chuzhouense sp. nov. is proposed. The type strain is DH-5T (=ACCC 19856T=KCTC 42746T).
Assuntos
Fazendas , Filogenia , Microbiologia do Solo , Sphingobacterium/classificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Sphingobacterium/genética , Sphingobacterium/isolamento & purificação , Vitamina K 2/análogos & derivados , Vitamina K 2/químicaRESUMO
A yellow-pigmented bacterial strain, designated Y2T, was isolated from farmland soil in Bengbu, Anhui province, China. Cells of strain Y2T were Gram-stain-negative, strictly aerobic, non-motile and rod-shaped. Strain Y2T grew optimally at pH 7.0, 30 °C and in the presence of 2 % (w/v) NaCl. The DNA G+C content was 68.9âmol%. The major fatty acids (>5 %) were iso-C15 : 0, iso-C17 : 0, summed feature 9 (C16 : 0 10-methyl and/or iso-C17 : 1ω9c), iso-C11 : 0 3-OH and iso-C11 : 0. The major respiratory quinone was ubiquinone-8 (Q-8), and the major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Phylogenetic analysis of the 16S rRNA gene sequences showed that strain Y2T was most closely related to Luteimonas mephitis B1953/27.1T (99.1 % 16S rRNA gene sequence similarity), followed by Luteimonas lutimaris G3T (98.6 %), Luteimonas abyssi XH031T (96.2 %) and Luteimonas aquatica RIB1-20T (96.0 %). Strain Y2T exhibited low DNA-DNA relatedness with Luteimonas mephitis B1953/27.1T (43.6 ± 0.5 %) and Luteimonas lutimaris G3T (43.9 ± 2.1 %). On the basis of phenotypic, genotypic and phylogenetic evidence, strain Y2T represents a novel species of the genus Luteimonas, for which the name Luteimonas soli sp. nov. is proposed. The type strain is Y2T ( = ACCC 19799T = KCTC 42441T).
Assuntos
Agricultura , Filogenia , Microbiologia do Solo , Xanthomonadaceae/classificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Fosfolipídeos/química , Pigmentação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/química , Xanthomonadaceae/genética , Xanthomonadaceae/isolamento & purificaçãoRESUMO
AIM: To study the transduction and localization mechanism of Marek's disease virus serotype 1 (MDV-1) CVI988 VP22 in different cells. METHODS: VP22 was expressed with recombinant adenovirus and identified by immunofluorescence assay (IFA) and Western blot. Lysates of recombinant virus infected 293 cells were added to normal MDBK cells to identify the transduction property of VP22. AD-293 cells infected with recombinant virus were fixed to investigate localization of VP22 at different time post infection. Transient expression of VP22 in AD-293 cells was carried out for control. RESULTS: The results showed that the VP22 expressed by recombinant adenovirus entered almost all the monolayer cells, which indicate the VP22 remains its transduction property. The VP22 first gather round the nucleus membrane, and then concentrated in particles in cytoplasm of 293 cells infected with recombinant adenovirus, compared with the nuclei localization pattern of VP22 in MDV infected CEF and transient expressed VP22 in 293 cells. CONCLUSION: The VP22 presented a different localization pattern in cells infected with different recombinant virus.
Assuntos
Adenoviridae/genética , Herpesvirus Galináceo 2/fisiologia , Doença de Marek/virologia , Proteínas Virais/metabolismo , Adenoviridae/metabolismo , Animais , Aves , Bovinos , Linhagem Celular , Citoplasma/virologia , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Herpesvirus Galináceo 2/genética , Humanos , Membrana Nuclear/virologia , Transporte Proteico , Proteínas Virais/genéticaRESUMO
AIM: To prepare and characterize monoclonal antibodies (mAb) against VP22 carboxyl terminus of CVI988/Rispens strain of Marek's disease virus serotype 1. METHODS: Carboxyl terminus of CVI988 VP22 (94aa-243aa) was expressed in prokaryotic system. mAbs against VP22 were prepared by hybridoma technology from BALB/c mice immunized with the fusion protein GST-VP22C and characterized by ELISA, indirect fluorescence assay (IFA) and Western blot. RESULTS: Two hybridoma cell lines stably secreting mAb against VP22C were obtained and designated as 3F7 and 4E4. mAb 3F7 could react with VP22 expressed in all the plaques, while mAb 4E4 stained all the cells nuclei in MDV-infected CEF cells. It was also found that 3F7 could react with VP22 expressed in Sf9 cells and denatured VP22 by Western blot analysis. In addition, it was further showed that the epitope of mAb 3F7 was located within the domain between 94aa and 193aa, the predicted site of protein transduction domain of VP22. CONCLUSION: The preparation of the mAb is very important to further research in protein transduction domain of MDV-1 VP22.