Abstract：
:Chronic infection of the hepatitis B virus (HBV) is one of the causes leading to liver cancer. The 3.2kb genome of HBV encodes four proteins: core antigen, surface antigen, a DNA polymerase and the X protein (HBx). The biological functions of HBx are not fully understood. It has been shown that HBx is a potent trans-activator, which activates transcription of many cellular and viral promoters indirectly via protein-protein interactions. These transactivating activities of HBx may contribute to the development of hepatocellular carcinoma. In this paper a truncated mini-HBx(-Cys) (18-142) protein, where the cysteines had been either deleted or substituted by serines, was constructed by site-directed mutagenesis and overexpressed as a 6xHis fusion protein in Escherichia coli. The 6xHis-mini-HBx(-Cys) protein was isolated from inclusion bodies, purified by Ni-affinity chromatography under denaturing conditions and refolded by sequential dialysis. The structure of the 6xHis-mini-HBx(-Cys) protein was analyzed by circular dichroism, fluorescence and one-dimensional NMR spectroscopic assays. The data presented here suggest that HBx is unstructured but has a propensity to gain secondary structure under specific experimental conditions. Its conformational flexibility might partially explain its functional complexity, namely its capacity to interact with a wide array of signaling proteins, transcriptional regulators and nucleic acids.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Rui E,Moura PR,Gonçalves Kde A,Kobarg Jdoi
10.1016/j.jviromet.2005.01.022subject
Has Abstractpub_date
2005-06-01 00:00:00pages
65-74issue
1-2eissn
0166-0934issn
1879-0984pii
S0166-0934(05)00037-6journal_volume
126pub_type
杂志文章abstract：:The first case of pandemic influenza A/H1N1/2009 infection was reported in Mexico in mid-April, 2009, and to date the new H1N1 virus has spread to over 160 countries. Therefore, it is important to obtain reliable epidemiological data on the spread of this virus by novel molecular methods can be developed in detection ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.11.033
更新日期：2010-05-01 00:00:00
abstract：:The GPRIME (Group PRIMEr design) programs examine aligned sets of gene sequences to discover homologous regions to be targeted in diagnostic tests. The core program moves a 'window' over the aligned sequences and calculates, at each window position, a 'redundancy value', namely the number of sequences that would repre...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00070-6
更新日期：1998-09-01 00:00:00
abstract：:A fluorogenic reverse transcriptase-polymerase chain reaction (RT-PCR) system was developed for use as a rapid diagnostic test for determining dengue viremia. The dengue virus 3'-noncoding sequence was utilized to formulate serotype-specific RT-PCR assays for quantitative identification of the four different dengue vi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(01)00280-4
更新日期：2001-06-01 00:00:00
abstract：:A bovine in vitro organ culture (BIVOC) system was evaluated as a model to study host and pathogen events during the course of bovine herpesvirus-1 infection. Upper respiratory tract epithelium, from slaughtered animals, was cultured in an air-liquid interface system and integrity, viability, and TNF-alpha gene expres...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.02.001
更新日期：2009-06-01 00:00:00
abstract：:A selected number of PCR protocols were evaluated to determine if they could serve as a universal protocol for detecting and identifying all arboviruses. In this study, four parameters that affect the efficacy of RT-PCR (RNA extraction method, choice of reverse transcriptase, choice of DNA polymerase and thermocycling...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00003-2
更新日期：1998-05-01 00:00:00
abstract：:Plum pox virus (PPV) causes the most destructive viral disease known as plum pox or Sharka disease in stone fruit trees. As an important regulated pathogen, detection of PPV is thus of critical importance to quarantine and eradication of the spreading disease. In this study, the innovative development of two AmplifyRP...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.06.026
更新日期：2014-10-01 00:00:00
abstract：:Two methods for rapid detection of adenovirus were tested: (i) 24-well plate centrifugation followed by staining with a monoclonal antibody after incubation for 24 h and 48 h, and (ii) pretreatment of A549 cells used in conventional cell culture and 24-well plate centrifugation with 10(-5)M dexamethasone. Twenty-seven...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(88)90144-9
更新日期：1988-06-01 00:00:00
abstract：:We applied real-time RT-PCR to the analysis of Tobacco rattle virus (TRV)-mediated virus-induced gene silencing (VIGS) of the phytoene desaturase (PDS) gene in Nicotiana benthamiana and tomato. Using a combination of direct measurement and mathematical assessment, we evaluated three plant genes, ubiquitin (ubi3), elon...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2006.07.017
更新日期：2006-12-01 00:00:00
abstract：:Mammalian orthoreoviruses (reoviruses) have served as highly useful models for studies of virus replication and pathogenesis. The development of a plasmid-based reverse genetics system represented a major breakthrough in reovirus research. The current reverse genetics systems for reoviruses rely on the expression of T...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.10.023
更新日期：2014-02-01 00:00:00
abstract：:In recent years, there has been an increase in reported cases of fowl adenovirus serotype 4 (FAdV-4) in chickens in China. The use of live attenuated vaccines contaminated with FAdV-4 has been proved to be one of the important causes of massive outbreaks of hydropericardium syndrome. To detect the contamination with F...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.09.005
更新日期：2019-03-01 00:00:00
abstract：:Bacterial ghosts offer a new avenue for the study of inactivated vaccines. However, for many years the mechanism of genetic inactivation was controversial. To obtain mouse monoclonal antibodies (mAbs) against protein E will allow the observation of its location and dynamic expression to expand understanding of the lys...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.03.007
更新日期：2013-05-01 00:00:00
abstract：:No effective prophylactic or therapeutic vaccine against HIV-1 in humans is currently available. This study analyzes the immunogenicity and safety of a recombinant attenuated vaccinia virus. A chimeric gene of HIV-1 multi-epitope genes containing CpG ODN and cholera toxin B subunit (CTB) was inserted into Chinese vacc...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.03.007
更新日期：2015-07-01 00:00:00
abstract：:The recent emergence of novel viruses requires reliable methodology for their identification and confirmation both on a cellular and molecular level. Mass spectrometry offers a suitable approach for the identification and characterisation of viral proteins and its application is demonstrated in this study. Menangle vi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(01)00441-4
更新日期：2002-04-01 00:00:00
abstract：:HIV-1 RNA viral load has become the major biological marker for disease prognosis and outcome of antiretroviral therapy in the treatment of HIV-infected individuals. The aim of this study was to compare the performance of the new CE marked NucliSens EasyQ HIV-1 assay with NucliSens HIV-1 QT assay (reference method). N...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.04.017
更新日期：2005-10-01 00:00:00
abstract：:Standard methods for assessing the thermal stability of viruses can be time consuming and rather qualitative yet such data is a necessary requisite for vaccine formulation. In this study a novel plate-based thermal scanning assay for virus particle stability has been developed (PaSTRy: Particle Stability Thermal Relea...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2012.06.014
更新日期：2012-10-01 00:00:00
abstract：:Green fluorescent protein (GFP) was targeted into bacteriophage T4 heads and proheads as a probe of the internal environment. Targeting was accomplished with internal protein III (IPIII) fusion proteins or capsid targeting sequence (CTS)-tagged proteins, where CTS is the 10-amino acid residue CTS of IPIII. Recombinant...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(00)00166-x
更新日期：2000-07-01 00:00:00
abstract：:The in vitro syncytium induction capacity of human immunodeficiency virus type 1 (HIV1) isolates is an important marker in the progression of the disease. Two methods have been widely used to determine the biological phenotype of HIV1. These two methods, the direct MT-2 assay and the supernatant assay, were compared f...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(96)02152-0
更新日期：1997-03-01 00:00:00
abstract：:In vitro studies related to various viral pathogenesis in swine have been hampered by the lack of relevant porcine cell lines. The susceptibility to porcine rotavirus infection was evaluated by using a newly established porcine intestinal epithelial cell line. Immunohistochemical staining for cytokeratin confirmed tha...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.03.007
更新日期：2014-06-01 00:00:00
abstract：:Cutting edge immune monitoring techniques increasingly measure multiple functional outputs for various cell types, such as intracellular cytokine staining (ICS) assays that measure cytokines expressed by T cells. To date, however, there is no precise method to measure virus-specific cytokine production by both T cells...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.01.003
更新日期：2014-04-01 00:00:00
abstract：:As a prelude to the development of nucleic acid probes specific for non-A, non-B hepatitis virus(es) (NANBV), plasmas with alanine aminotransferase levels greater than or equal to 110 IU were assayed for DNA by a radioimmunoassay. Approximately 50% of such plasmas are expected to contain NANBV. One-hundred and seventy...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(84)90042-9
更新日期：1984-02-01 00:00:00
abstract：:We developed a non-radioisotopic (non-RI) reverse transcriptase assay (RTA). The reverse transcriptase (RT) incorporates biotin-11-deoxyuridine-triphosphate (bio-dUTP) using a poly(rA) template hybridized with oligo(dT) primer that is immobilized on the surface of a 96-well microtiter plate. This assay is thus semi-au...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(92)90063-j
更新日期：1992-11-01 00:00:00
abstract：:Water is one major pathways by which the white spot syndrome virus (WSSV) pathogen enters aquaculture facilities. This paper describes the production and use of a capacitive biosensor for the quantitative detection of as little as 1copy/μl of WSSV in shrimp pond water. A glutathione-S-transferase tag for white spot bi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.01.010
更新日期：2011-04-01 00:00:00
abstract：:Asthma and chronic obstructive pulmonary disease exacerbations are associated with human rhinovirus (HRV) lung infections for which there are no current effective antiviral therapies. To date, HRV infectivity of cells in vitro has been measured by a variety of biochemical and immunological methods. This paper describe...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.02.002
更新日期：2011-05-01 00:00:00
abstract：:This paper describes the validation of a solid-phase competition enzyme-linked immunosorbent assay (SPCE) for the serological detection of antibody to serotype O foot-and-mouth disease (FMD) in sheep, cattle and pigs. The specificity of the SPCE was calculated from the results of testing known negative sera from sheep...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2003.09.016
更新日期：2004-02-01 00:00:00
abstract：:Rotavirus (RoV) and Norovirus (NoV) are the main causes of viral gastroenteritis. Currently, there is no validated multiplex real-time PCR that can detect and quantify RoV and NoV simultaneously. The aim of the study was to develop, validate, and internally control a multiplex one-step RT real-time PCR to detect and q...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2012.09.021
更新日期：2013-01-01 00:00:00
abstract：:Arboviruses in the genus Flavivirus are major causes of human disease worldwide, and yet they are widely under reported. This is partially due to the classical methods of detection, using virologic or immunologic techniques, which typically lack sensitivity and specificity or require biological containment facilities ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2007.05.001
更新日期：2007-10-01 00:00:00
abstract：:Indicator cell lines are useful biological tools for monitoring virus infection. In order to monitor infection with bovine immunodeficiency virus (BIV) in vitro, an indicator cell line derived from baby hamster kidney cells which contains integrated copies of an enhanced green fluorescent protein gene driven by the BI...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.07.007
更新日期：2010-01-01 00:00:00
abstract：:Rabies virus is a neurotropic agent which spreads in the CNS via axonal transport. Previous studies had shown that this axonal transport through the brain could be inhibited by stereotaxic administration of colchicine; however, this inhibition was reversible. We describe here a method to enhance the duration of this c...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(90)90089-x
更新日期：1990-04-01 00:00:00
abstract：:With the approval of the first HIV-1 integrase inhibitor raltegravir and a second one in phase III clinical development (elvitegravir), genotypic and phenotypic resistance assays are required to guide antiretroviral therapy and to investigate treatment failure. In this study, a genotypic and phenotypic recombinant vir...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.06.015
更新日期：2009-11-01 00:00:00
abstract：:Treatment of chronic hepatitis C is associated with varying success rates, substantial medical costs and serious side effects. Several host polymorphisms have been identified near the IL28B gene, of which the homozygous rs12979860 CC was found to be associated with significantly favourable treatment outcome. To determ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.04.018
更新日期：2011-07-01 00:00:00
