RESUMO
NSscon (23 aa), a common epitope in the gene silencing suppressor NSs proteins of the members of the Watermelon silver mottle virus (WSMoV) serogroup, was previously identified. In this investigation, we expressed different green fluorescent protein (GFP)-fused deletions of NSscon in bacteria and reacted with NSscon monoclonal antibody (MAb). Our results indicated that the core 9 amino acids, "(109)KFTMHNQIF(117)", denoted as "nss", retain the reactivity of NSscon. In bacterial pET system, four different recombinant proteins labeled with nss, either at N- or C-extremes, were readily detectable without position effects, with sensitivity superior to that for the polyhistidine-tag. When the nss-tagged Zucchini yellow mosaic virus (ZYMV) helper component-protease (HC-Pro) and WSMoV nucleocapsid protein were transiently expressed by agroinfiltration in tobacco, they were readily detectable and the tag's possible efficacy for gene silencing suppression was not noticed. Co-immunoprecipitation of nss-tagged and non-tagged proteins expressed from bacteria confirmed the interaction of potyviral HC-Pro and coat protein. Thus, we conclude that this novel nss sequence is highly valuable for tagging recombinant proteins in both bacterial and plant expression systems.
Assuntos
Epitopos/metabolismo , Proteínas Recombinantes de Fusão/isolamento & purificação , Proteínas Recombinantes de Fusão/metabolismo , Tospovirus/genética , Proteínas não Estruturais Virais/metabolismo , Sequência de Aminoácidos , Biotecnologia , Western Blotting , Cucurbita/genética , Epitopos/química , Epitopos/imunologia , Escherichia coli/genética , Proteínas de Fluorescência Verde , Imunoprecipitação , Dados de Sequência Molecular , Vírus do Mosaico/genética , Oligopeptídeos/química , Oligopeptídeos/genética , Oligopeptídeos/metabolismo , Folhas de Planta/química , Folhas de Planta/metabolismo , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas não Estruturais Virais/química , Proteínas não Estruturais Virais/genética , Proteínas não Estruturais Virais/imunologiaRESUMO
OBJECTIVE: The aim of this study was to scrutinize the literature to determine the efficacy and safety of gamma knife surgery (GKS) for the treatment of nonfunctioning pituitary adenomas (NFPAs) with volumetric classification. METHODS: Electronic databases including MedLine, PubMed, and Cochrane Central were searched. The literature related to patients with NFPAs treated with GKS was collected. Eligible studies reported on the rate of tumor control (RTC), the rate of radiosurgery-induced optic neuropathy injury (RRIONI), the rate of radiosurgery-induced endocrinological deficits (RRIED), and other parameters. RESULTS: A total of 17 studies met the criteria. based on the tumor volume, nfpas were divided into three groups: the RTC of group I (93 patients) with tumor volumes <2 ml was 99% (95% CI 96-100%), the RRIONI was 1% (95% CI 0-4%), and the RRIED was 1% (95% CI 0-4%). The RTC of group II (301 patients) with volumes from 2 to 4 ml was 96% (95% CI 92-99%), the RRIONI was 0 (95% CI 0-2%), and RRIED was 7% (95% CI 2-14%). The RTC of group III (531 patients) with volumes larger than 4 ml was 91% (95% CI 89-94%), the RRIONI was 2% (95% CI 0-5%), and the RRIED was 22% (95% CI 14-31%). There were significant differences in the RTC and in the RRIED among the three groups (P<0.001), indicating that there were higher RRIED and lower RTC with the increase of tumor volume. CONCLUSIONS: NFPAs, according to tumor volume classification, need stratification for GKS treatment. GKS is the optimal choice for the treatment of group II NFPAs. Patients with residual tumor volumes of <4 ml will benefit most from GKS treatment.