Journal: Biochemical and Biophysical Research Communications

Article Title: The SARS-CoV S glycoprotein: expression and functional characterization

doi: 10.1016/j.bbrc.2003.11.054

Figure Lengend Snippet: Cell fusion mediated by the S glycoprotein. (A) Syncytium formation between 293T cells transfetced with pSecTag2B-S and pCDNA3-ACE2, respectively (right panel). There was no syncytium formation between 293T cells transfected with pSecTag2B-S and pCDNA3-ACE2-Ecto (left panel). (B) Cell fusion measured by a reporter gene-based assay. S glycoprotein expressed in both pCDNA3 and pSecTag2B vectors can be used in a β-gal reporter gene-based cell–cell fusion assay. A pCDNA3-based plasmid without S insert was used as plasmid control, and fusion between S-expressing cells with ACE2-ecto expressing cells was used as negative control.

Article Snippet: The fragment was cut with Bam HI and Apa I and cloned into corresponding sites of expression vector pSecTag2B (Invitrogen).

Techniques: Transfection, Reporter Gene Assay, Cell-Cell Fusion Assay, Plasmid Preparation, Expressing, Negative Control

Journal:

Article Title: Specific Antibodies to Porphyromonas gingivalis Lys-Gingipain by DNA Vaccination Inhibit Bacterial Binding to Hemoglobin and Protect Mice from Infection

doi: 10.1128/IAI.69.5.2972-2979.2001

Figure Lengend Snippet: Construction of plasmid DNA for immunization. Amplified kgpcd and rgpcd genes were inserted into the eukaryotic expression vectors pSecTag2A and pSecTag2B, respectively (see Materials and Methods). SV40, simian virus 40; PCMV, human cytomegalovirus immediate-early promoter; BGH, bovine growth hormone gene (provides the polyadenylation [pA] signal); Ampr, ampicillin resistance gene; Zeor, Zeocin resistance gene.

Article Snippet: To generate the rRGP cd polypeptide, the rgp cd fragments used for pSecTag2B/ rgp cd construction were inserted into pGEX2T (Amersham Pharmacia Biotech), which is the vector for expression of the glutathione S -transferase (GST) fusion protein.

Techniques: Plasmid Preparation, Amplification, Expressing

Journal:

Article Title: Specific Antibodies to Porphyromonas gingivalis Lys-Gingipain by DNA Vaccination Inhibit Bacterial Binding to Hemoglobin and Protect Mice from Infection

doi: 10.1128/IAI.69.5.2972-2979.2001

Figure Lengend Snippet: Induction of IgG antibodies against rKGPcd and rRGPcd in serum from DNA-immunized mice. Mice were immunized by injection of pSecTag2A/kgpcd (50 μg) and pSecTag2B/rgpcd (50 μg), respectively. At 0, 7, 14, 21, 28, and 35 days after the primary immunization, sera were collected to evaluate specific antibody responses to rKGPcd (A) and rRGPcd (B). Control mice were immunized with intact pSecTag2A plasmid (50 μg) or PBS (50 μl). The values are expressed as means and standard deviations of log2 ELISA antibody titers.

Article Snippet: To generate the rRGP cd polypeptide, the rgp cd fragments used for pSecTag2B/ rgp cd construction were inserted into pGEX2T (Amersham Pharmacia Biotech), which is the vector for expression of the glutathione S -transferase (GST) fusion protein.

Techniques: Injection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Specific Antibodies to Porphyromonas gingivalis Lys-Gingipain by DNA Vaccination Inhibit Bacterial Binding to Hemoglobin and Protect Mice from Infection

doi: 10.1128/IAI.69.5.2972-2979.2001

Figure Lengend Snippet: Western blot analysis of serum samples. Immunoblot analyses of serum samples from mice immunized by plasmid DNA are shown. (A) Profiles probed with serum elicited by pSecTag2A/kgpcd. (B) Profiles probed with serum elicited by pSecTag2B/rgpcd. Lanes: a, whole-cell extracts of P. gingivalis ATCC 33277 (40 μg); b, whole-cell extracts of P. gingivalis W50 (40 μg); c, rKGPcd (2 μg); d, rRGPcd (2 μg).

Article Snippet: To generate the rRGP cd polypeptide, the rgp cd fragments used for pSecTag2B/ rgp cd construction were inserted into pGEX2T (Amersham Pharmacia Biotech), which is the vector for expression of the glutathione S -transferase (GST) fusion protein.

Techniques: Western Blot, Plasmid Preparation

Journal:

Article Title: Specific Antibodies to Porphyromonas gingivalis Lys-Gingipain by DNA Vaccination Inhibit Bacterial Binding to Hemoglobin and Protect Mice from Infection

doi: 10.1128/IAI.69.5.2972-2979.2001

Figure Lengend Snippet: Disease status of P. gingivalis infection in immunized mice

Article Snippet: To generate the rRGP cd polypeptide, the rgp cd fragments used for pSecTag2B/ rgp cd construction were inserted into pGEX2T (Amersham Pharmacia Biotech), which is the vector for expression of the glutathione S -transferase (GST) fusion protein.

Techniques: Infection