Part:BBa_K4875007

Modified core streptavidin

Contribution by 2023 iGEM Team Metropolis-Shanghai

Basic Part: BBa_K4875007

Group: Metropolis-Shanghai iGEM 2023

Profile

Name: Modified core streptavidin

Base Pairs: 600 bp

Origin: Streptomyces avidinii, Synthetic

Properties

Usage and Biology

Streptavidin is a protein secreted by Streptomyces avidinii^[1], with a molecular weight of 66kD. Modified core streptavidin protein has a molecular weight of 21.6 kD. Each peptide chain can bind to one biotin, that is, one streptavidin can bind to four biotin molecules^[2].

Application: There are various forms of application in ELISA

1. Used for indirect packaging
2. Used to amplify detection signals and improve detection sensitivity

In our project, an amino acid fragment (KDEL) residing in the endoplasmic reticulum was used as the hook protein, which was connected to Streptavidin (Str)(BBa_K4875007). In addition, α2A-AR was connected to Streptavidin binding protein (SBP)(BBa_K4875012) and Rluc8(BBa_K4875014), and the genes of these two components were integrated into a plasmid for co-expression. This way, the newly generated protein would be left in the endoplasmic reticulum and stopped further transport to the cell membrane. As shown in Figure 1.

Figure 1. Schematic diagram of RUSH-BRET assay. Image by Xu Xing.^[3]

References:

1. Chaiet L, Wolf FJ. The properties of streptavidin, a biotin-binding protein produced by Streptomycetes. Arch. Biochem. Biophys. 1964;106:1–5.
2. Green NM. Avidin and streptavidin. Methods Enzymol. 1990;184:51–67.
3. Xu X, Wu G. Human C1orf27 protein interacts with α2A-adrenergic receptor and regulates its anterograde transport. J Biol Chem. 2022 Jun;298(6):102021.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]