Promoter (J23100, Anderson Strong)

Constitutive promoter family

Parts J23100 through J23119 are a family of constitutive promoter parts isolated from a small combinatorial library. J23119 is the "consensus" promoter sequence and the strongest member of the family. These promoter parts can be used to tune the expression level of constitutively expressed parts.

RBS (B0034, Ribosome Binding Site)

RBS based on Elowitz repressilator.

Designed by: Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.

Terminator (B0010)

Transcriptional terminator consisting of a 64 bp stem-loop.

Designed by: Randy Rettberg

Terminator (B0012)

Transcription terminator for the E. coli RNA polymerase.

Designed by: Reshma Shetty

GBP (K2885001, Gold-Binding Polypeptide)

http://parts.igem.org/Part:BBa_K2885001

Gold binding polypeptide (GBP) can be used as an anchor component, robustly binding to a gold surface and it is consisted of 14 amino acids (MHGKTQATSGTIOS) triplication. The strong coupling property of GBP onto a gold-patterned substrate was delineated in a previous investigation. The binding peculiarity of GBP can be an useful avenue for protein immobilization on gold substrate, substantially contributing to the biosensor development.

ProG (K2885000, Protein G)

http://parts.igem.org/Part:BBa_K2885000

Protein G (ProG), one of the immunoglobulin-binding proteins, was derived from two streptococcal strains (group C and G) and has been used for purification of antibodies due to its binding characteristic to antibodies’ Fab and Fc region. Our ProG recombinant from Escherichia coli was used for efficient immobilization of antibodies, which enables us to develop our electrochemical biosensor.

GBP-ProG (K2885002, Gold Binding Protein, Protein G Fusion Protein)

http://parts.igem.org/Part:BBa_K2885002

In our study, gold binding polypeptide (GBP)-Protein G (ProG)(GBP-ProG), BBa_K2885002, was constructed from Integrated DNA Technologies (IDT) and the sequence was analyzed by BIOFACT™ sequencing analysis service. After the construct was inserted into pSB1C3 vector, the recombinant vector’s transformation and amplification were perfromed in E. coli DH5-alpha competent cell with IPTG-induced expression. With the TALON metal affinity resin, the GBP-ProG fusion protein was highly purified. The purified protein was confirmed by SDS-PAGE and then quantified using the Bradford protein assay to dilute to 1 mg/ml with a phosphate buffered solution (PBS) for experimental use in this study. We clarified that our GBP-ProG can be an efficient crosslinker of diverse antibodies.

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