Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle-5

Sence of chimeric GvpC proteins. Electroblotted wild type gas vesicles samples (wt-GV) prepared from the NRC-1 were incubated with purified rabbit anti-GvpC antibody or with monkey anti-SHIV antibody. As shown, the GvpC protein is clearly detected by anti-GvpC antibody (), but this protein is not detected by anti-SHIV antibody (). In cell lysates, Tat and Rev sequences contained in the putative recombinant GvpC protein are each detected using anti-SHIV antibody (plasma number R94085) from a monkey infected with SHIV virus (). The recombinant chimeric protein bands identified here correspond in size to bands identified with anti-GvpC antibody (see ).<p><b>Copyright information:</b></p><p>Taken from "Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle"</p><p>http://www.biomedcentral.com/1472-6750/8/9</p><p>BMC Biotechnology 2008;8():9-9.</p><p>Published online 31 Jan 2008</p><p>PMCID:PMC2270826.</p><p></p

Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle-8

Ments of sera taken at week 43 demonstrate that notable titers remain at this very late time point: 1:160 for Rev; and for Tat and Nef1, the titers are ≥ 1:1,280. : for this graph the absorbance scale is 0.0 – 5.0 and note also that indicates only one animal remained in the Rev group at 43 weeks.<p><b>Copyright information:</b></p><p>Taken from "Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle"</p><p>http://www.biomedcentral.com/1472-6750/8/9</p><p>BMC Biotechnology 2008;8():9-9.</p><p>Published online 31 Jan 2008</p><p>PMCID:PMC2270826.</p><p></p

Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle-0

Thin the gene, allowed insertion of the exogenous DNA fragments. The unique restriction enzyme sites Spe I, AsiS I and Afl II allowed manipulation/isolation of this cluster of gas vesicle genes by appropriate restriction digests. The antibiotic resistance genes are indicated with the boxes and allow selection of transformed or SD109 using Ampicilin (Amp) and Mevinolin (mev), respectively.<p><b>Copyright information:</b></p><p>Taken from "Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle"</p><p>http://www.biomedcentral.com/1472-6750/8/9</p><p>BMC Biotechnology 2008;8():9-9.</p><p>Published online 31 Jan 2008</p><p>PMCID:PMC2270826.</p><p></p

Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle-10

He recombinant genes. The agarose gel shows the PCR amplicon sizes of the selected SIV gene fragments and are indicated along the horizontal axis: :150, 243 and :642 bp. Their presence inherently verifies SIV DNA retention by each r-genes. The Control (C) is an internal fragment from the gene and serves as a marker for the migration of this 350 bp fragment. The relevant primers are shown in Table 1.<p><b>Copyright information:</b></p><p>Taken from "Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle"</p><p>http://www.biomedcentral.com/1472-6750/8/9</p><p>BMC Biotechnology 2008;8():9-9.</p><p>Published online 31 Jan 2008</p><p>PMCID:PMC2270826.</p><p></p

Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle-2

or carrying the native leftward operon of the gas vesicle gene cluster and the rightward operon carrying an inserted SIV fragment of interest. Original magnifications 1,000 ×.<p><b>Copyright information:</b></p><p>Taken from "Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle"</p><p>http://www.biomedcentral.com/1472-6750/8/9</p><p>BMC Biotechnology 2008;8():9-9.</p><p>Published online 31 Jan 2008</p><p>PMCID:PMC2270826.</p><p></p

Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle-9

Thin the gene, allowed insertion of the exogenous DNA fragments. The unique restriction enzyme sites Spe I, AsiS I and Afl II allowed manipulation/isolation of this cluster of gas vesicle genes by appropriate restriction digests. The antibiotic resistance genes are indicated with the boxes and allow selection of transformed or SD109 using Ampicilin (Amp) and Mevinolin (mev), respectively.<p><b>Copyright information:</b></p><p>Taken from "Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle"</p><p>http://www.biomedcentral.com/1472-6750/8/9</p><p>BMC Biotechnology 2008;8():9-9.</p><p>Published online 31 Jan 2008</p><p>PMCID:PMC2270826.</p><p></p

Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle-7

S display the temporal profiles of antigen specific antibody responses to the SIV peptide expressed within the GvpC protein. For Tat, all post-4 week samples exhibit a titer of ≥ 1:1,280 while for Rev, the maximal titer, 1:320 occurs in the week 2 post second booster sera. For Nef1, the highest titer, 1:320, is evident in sera from week 8 post second booster and by the 12 week time point, only sera from the animals immunized with r-GV expressing Rev retained a peptide specific titer.<p><b>Copyright information:</b></p><p>Taken from "Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle"</p><p>http://www.biomedcentral.com/1472-6750/8/9</p><p>BMC Biotechnology 2008;8():9-9.</p><p>Published online 31 Jan 2008</p><p>PMCID:PMC2270826.</p><p></p

Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle-1

The recombinant genes. The agarose gel shows the PCR amplicon sizes of the selected SIV gene fragments and are indicated along the horizontal axis: :150, 243 and :642 bp. Their presence inherently verifies SIV DNA retention by each r-genes. The Control (C) is an internal fragment from the gene and serves as a marker for the migration of this 350 bp fragment. The relevant primers are shown in Table 1.<p><b>Copyright information:</b></p><p>Taken from "Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle"</p><p>http://www.biomedcentral.com/1472-6750/8/9</p><p>BMC Biotechnology 2008;8():9-9.</p><p>Published online 31 Jan 2008</p><p>PMCID:PMC2270826.</p><p></p

Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle-4

samples are resolved in 8–16% SDS-PAGE gels, immunoblotted and probed with rabbit anti-GvpC antibody. The size range of the protein standards (kDa) is indicated at the left and the recombinant GvpC protein bands are identified at the right.<p><b>Copyright information:</b></p><p>Taken from "Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle"</p><p>http://www.biomedcentral.com/1472-6750/8/9</p><p>BMC Biotechnology 2008;8():9-9.</p><p>Published online 31 Jan 2008</p><p>PMCID:PMC2270826.</p><p></p

Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle-6

. The Coomassie Blue stained 12% SDS-PAGE gels shows resolution of proteins in gas vesicles or lysates from these transformed SD109 cultures (). Duplicates of these resolved proteins were electroblotted () and the presence of the Nef1 peptide in chimeric r-GvpC, identified by both absorbed anti-Nef1 sera from mice immunized with r-GVand the monkey anti-SHIV antibody (plasma number R94085). Antibody binding shows recognition of Nef1 peptide within the recombinant GvpC protein. The mouse anti-Nef1 serum was used at a 1:250 dilution; the monkey anti-SHIV antibody was used at a 1:200 dilution. The standards (kDa) are shown on the left and the immuno detected band for recombinant Nef1 containing r-GvpC protein is identified on the right.<p><b>Copyright information:</b></p><p>Taken from "Recombinant gas vesicles from . displaying SIV peptides demonstrate biotechnology potential as a pathogen peptide delivery vehicle"</p><p>http://www.biomedcentral.com/1472-6750/8/9</p><p>BMC Biotechnology 2008;8():9-9.</p><p>Published online 31 Jan 2008</p><p>PMCID:PMC2270826.</p><p></p