GTA MCP | Gene Transfer Agent (GTA) major capsid protein (MCP)
AS08 365 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Rhodobacterales
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KLH-conjugated conserved peptide sequence found in the Gene Transfer Agent (GTA) major capsid protein (MCP) encoded in Bacteria within the Rhodobacterales order of the class alpha-Proteobacteria including Rhodobacter sphaeroides UniProt: Q3J3K4
31.4 | 32 kDa (predicted mature capsid protein of Rhodobacter capsulatus)
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Roseobacter capsulatus cells, pelleted by centrifugation and resuspended in an equial volume of TE buffer and supernatant sample* were separated on 10% SDS-PAGE and blotted 1h to nitrocellulose. The "+" indicated R. capsulatus SB1003 (GTA positive) and "-" indicated R. capsulatus A1 (GTA capsid protein negative). Blots were blocked in 5% skim milk in TBST followed by incubation with anti-GTA antibodies (AS08 365) at dilution 1: 1000 at 4°C over night. After washes blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Santa Cruz Biotechnology, Santa Cruz, CA) and specific bands were detected with chemiluminescence detection reagent.Exposure time was 30 seconds with CCD camera.
* - supernatant sample was obtained in a following way: cells were removed by two rounds of centrifugation at 17,000 g for 2 min. with sub-sample of the supernatant removed to a new tube. Two volumes of the cells or final culture supernatant were mixed with 1 volume of 3X SDS-PAGE sample buuffer (NEB).
Gene Transfer Agents (GTAs) are bacteriophage-like particles which function as a means to package and transfer genomic DNA between bacterial cells.GTA-mediated gene transfer might be an important phenomenon in natural microbial communities.
Tomasch et al. (2018). Packaging of Dinoroseobacter shibae DNA into Gene Transfer Agent Particles Is Not Random. Genome Biol Evol. 2018 Jan 1;10(1):359-369. doi: 10.1093/gbe/evy005.
Mercer and Lang (2014). Identification of a predicted partner-switching system that affects production of the gene transfer agent RcGTA and stationary phase viability in Rhodobacter capsulatus. BMC Microbiol. 2014 Mar 19;14(1):71.
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