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Legionella pneumophila is an intracellular bacteria, and its successful parasitism in host cells involves two reciprocal phases: transmission and intracellular replication. In this study, we try to construct expression vectors of pcDNA3.1 (+)-lvgA/ctxB-EGFP fused gene and to investigate its transient expressions in NIH3T3 cells. Then preliminary analysis the immunogenicity of lvgA/ctxB fusion gene...
To clone the cDNA of mouse 14–3–3θ protein gene, investigate its prokaryotic expression and produce purified recombinant mouse 14–3–3θ protein. The recombinant plasmid containing mouse 14–3–3θ gene was transformed into E. Coli BL21 to express a fusion protein with mouse 14–3–3θ protein tagged by glutathione-S-transferase (GST). The fusion protein was then cleaved by thrombin during passing through...
In this study, the antigenic site B/C of S gene of transmissible gastroenteritis virus (TGEV) was cloned into prokaryotic expression vector pET-28a and the recombinant plasmid was transferred into E.coli BL21 (DE3). The fusion protein was expressed after induction with IPTG. The SDS-PAGE purified protein was used as immunogen. Six-week-old BALB/c mice were immunized three times with recombinant protein...
For raising the antigenicity of Mycobacterium bovis single antigen, fusion protein of two genes was acquired. The DNA fragments of mpb83 and ag85b were fused by splicing by overlapping extension (SOE) polymerase chain reaction(PCR), and the fusion gene mpb83-ag85b were cloned into pMD18-T vector, then we got the recombinant plasmid pMD-83-85b. pMD-83-85b and pET28a(+) were digested by BamH I and EcoR...
Lipoprotein lipase (LPL) of Tianfu meat goose was cloned and sequenced using a RT-PCR approach. The nucleotide sequence covered 468 bp with an open reading frame of 155 amino acids. Alignment analysis indicated that the nucleotide sequences are highly conserved to other species studied including chicken, domestic guinea pig, house mouse, Norway rat, cattle, cow and goat, and the homologies are 92...
We investigate the effects of anti-Fas ribozyme on Fas expression and apoptosis in primary cultured Neonatal mouse ventricular myocytes. Neonatal mouse ventricular myocytes were isolated by using collagenase irrigation. A hammerhead ribozyme targeting the Fas mRNA was constructed, and transfected into Neonatal mouse ventricular myocytes via Effectene. Then Fas expression in Neonatal mouse ventricular...
We present a global clustering approach to identify putative intergenic non-coding RNAs based on the RNA polymerase II and Histone 3 lysine 4 trimethylation signatures. Both of these signatures are processed from the digital sequencing tags produced by chromatin immunoprecipitation, a high-throughput massively parallel sequencing (ChIP-Seq) technology. Our method compares favorably to the comparison...
SET and MYND domain-containing protein 3 (SMYD3) is a novel histone methyltransferase involved in human carcinogenesis. To further establish the transgenic mice of SMYD3 and provide a useful platform for the study of SMYD3 and the screening of novel antitumor drugs, a transgene for liver-specific simultaneous expression of SMYD3 and EGFP was constructed. The human SMYD3 gene was subcloned into pIRES2-EGFP...
A phosphorylatable short peptide(pSP) with the amino acid composition of ??LLLRRRDNEYFYVRRLL?? containing two potentially phosphorylatable tyrosine residues was chemically synthesized. This short peptide was characteristic with the isoelectric point of 10.88 and could be positively charged under physiological condition. The ??DNEYFYV?? motif in the pSP is the substrate of Jak2 kinase which was verified...
A regenerable flow-through sensor based on white light reflectance spectroscopy for label-free detection of proteins and DNA is presented. Detection is based on monitoring the interference wavelength shift caused by an increase in film thickness during bioreactions. By employing an anti-mouse IgG antibody/mouse IgG assay, a detection sensitivity of 150 pM was determined. In addition, the sensor response...
To illustrate the possible function of genotoxicity of BBP on DNA, this study took mice livers as experimental materials to detect the levels of DNA-protein crosslinks (DPC), the activity of surperoxide dismutase (SOD), and the contents of malondialdehyde (MDA) with intraperitioneal injection of different doses of BBP for two consecutive 14 days. DPC levels in hepatic cells showed a dose-dependent...
Heroin addiction caused oxidative stress was testified for the first time in mice. After administered with heroin, mice not only showed decreased total antioxidant capacity as well as antioxidant enzymes such as, SOD, CAT and GPx in brain, but also include oxidative damage of DNA, protein and lipid. On the other hand, exogenous antioxidants could restrain the oxidative stress, even alleviate withdrawal...
The objective of this study is to explore the relation between oxidative stress and DNA-protein crosslink formation caused by formaldehyde (FA) inhalation in mice liver. 15 Kun Ming mice were divided into 3 groups: control group, 3.0 mg/m3 gaseous FA inhaled group, 3.0 mg/m3 gaseous FA and lipoic acid (LA) treated group. The content of reduced glutathione (GSH) and the content of DPC were measured...
Antibiotic resistance is a rapidly evolving problem that is not being adequately met by new antimicrobial drugs. Thus, there is a pressing need for effective antibacterial therapies that can be adapted against antibiotic-resistant bacteria. Here, we engineered synthetic bacteriophage to combat antibiotic-resistant bacteria by overexpressing proteins and attacking gene networks which are not directly...
Studies were conducted to establish the biochemical and pharmacological effects of W-2721 after irradiation in the mouse tests for up to 30 days of irradiation. Adult mice were irradiated with 3, 6, and 8 Gy of /sup 60/Co gamma rays with and without pretreatment by WR-2721 at a dose rate of 400 mg/kg body weight. Animals were sacrificed at different postirradiation days to estimate the amount of DNA,...
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