Using LANL GFP to Determine Solubility

Overview

LANL's GFP can be used to quickly determine whether an expressed protein is soluble. It does this very simply and elegantly. First, a short section of GFP called a "tether" or "tag" is appended onto the nucleotide sequence of the protein of interest. The tether is about 14 amino acids (a.k.a. S11, strand 11 of GFP) which is too short to effect the dynamics of your expressed protein.

Figure 1. Split GFP: how a soluble expressed protein will behave in the system.

A vector containing the remaining portion of GFP (a.k.a S1-10, strand 1-10 of GFP) called the "detector" is also inserted into the host cell. Once the target protein and S11, and the remaining portion of GFP, S1-10 are inserted into the host, determining the solubility of the target protein is easy: express your protein of interest (which has the S11 tag) (Fig. 1, #1), followed by inducing the expression of the S1-10 "detector" (Fig. 1, #2).

Figure 2. Split GFP: how an insoluble expressed protein will behave in the system.

If your protein is soluble, aggregation will not occur (Fig. 2, #1) and the S1-10 detector will be unable to bind to the S11 tag to create a fully functioning, fluorescing GFP (Fig. 1, #3). However, if the expressed protein is not soluble, it will aggregate (Fig. 2, #1), not allowing the S11 tag to interact with the S1-10 detector and no fluorescense will occur (Fig. 2, #3).

Key Characteristics

• Fast
• Cheap
• Reliable
• Quantifiable
  

To learn more, read about published results in our Technical Library.

RELATED LINKS

• Technical Library (pubs and presentations)
• GFP Wiki that defines common terms
• Feedback from our users