Recombinant HIV gag protein(Pro133-Leu363), His-tagged

• Cat.No.: gag-314H
• Product Overview: Recombinant HIV-1 gag protein (ACI05538.1)(Pro 133-Leu 363), known as core nucleocapsid protein P24 was expressed in E. coli, with a polyhistidine tag at the C-terminus and a Met at the N-terminus.

Specification

• Species: HIV
• Source: E.coli
• Tag: His
• Protein Length: Pro133-Leu363
• Form: Lyophilized from sterile 0.14M PB, 0.5M NaCl, 20% glycerol, 1mM DTT, pH 8.0. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween80 are added as protectants before lyophilization.
• Molecular Mass: The recombinant human HIV-1 Gag-p24 protein comprises 240 amino acids and has a predicted molecular mass of 26.5 kDa as estimated in SDS-PAGE under reducing conditions.
• Purity: > 95 % as determined by SDS-PAGE
• Storage: Samples are stable for up to twelve months from date of receipt at -20°C to -80°C. Store it under sterile conditions at -20°C to -80°C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.

Case Study

Case 1: Boix-Besora A, et al. Vaccines (Basel). 2022

VLPs are pretty cool because they act like real viruses without the harmful genetic material. This makes them great for developing vaccines. For instance, HIV-1 Gag VLPs have traits similar to the actual COVID-19 virus, which means we can use them as a base to create COVID-19 vaccines. In this study, researchers mixed HIV-1 Gag protein with coronavirus spike protein to make these VLPs. They tinkered with the process using a methodical approach to get it just right, and then we scaled it up for mass production, ensuring everything meets the industry standards. This technique isn't just for COVID-19; it has the potential to be adapted for other diseases too.

Fig1. Western blot of purified S-VLPs and G-VLPs.

Fig2. Transfection kinetics: percentage of Gag::eGFP-expressing cells at different time points after transfection.

Case 2: McKinstry WJ, et al. Protein Expr Purif. 2014

Hiv-1 Gag protein Pr55(Gag) is a multifunctional protein that plays an important role in HIV assembly. The structure of HIV-1 is known to be complex, so researchers have been trying to figure out the specific structure of Pr55(Gag). But the trouble is, these things tend to stick together, so there's not much you can extract. So this protein is extremely difficult to study. Fortunately, however, a study using a bacterial expression system successfully produced a recombinant version of HIV-1 Pr55(Gag). This time, they added a His×6 label, and used some professional separation methods, and finally made a batch of high-purity HIV-1 Pr55(Gag), and these proteins in solution behaved like trimers.

Fig1. SDS–PAGE analysis o fbacterial lysates.

Fig2. Analysis of IMAC purified HIV-1 Pr55Gag.

Application

The HIV gag protein is pretty important when it comes to making HIV vaccines. Why? Well, it's a key part of the virus's structure and plays a big role in how the virus goes through its life cycle, like piecing together new virus particles. Because it can kickstart the body's immune reaction to fight off HIV, it's been a hot topic for researchers. In vaccines, scientists tweak the gag protein to create something called virus-like particles, or VLPs. These look like the actual virus but don't have the genes that can make you sick. This makes them great for teaching the immune system how to react. The gag protein can even make VLPs on its own when put together in different systems, which is super useful for showing HIV-1 Env to the immune system.

The gag protein is a big deal in science and industry circles. In research, it helps scientists figure out how HIV puts itself together and makes copies, which is huge for coming up with better treatments. By diving deeper into this protein, researchers get a handle on how HIV is put together, which is crucial when working on vaccines and other treatments. On the industrial side, it's used in making vaccines that can prompt a strong immune response. It's also handy in tests for HIV, making them more reliable. All in all, the recombinant HIV gag protein has a lot of uses, from helping out in the lab to being part of clinical tools, playing a big part in the fight against HIV.

Fig1. A model that accounts for specific localization of Gag to the plasma membrane. (Christopher Sumner, 2022)