Scar

Part:BBa_K5384002:Design

Designed by: Si Cheng   Group: iGEM24_HBUT-Wuhan   (2024-08-23)


DDDDK


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]


Design Notes

The greatest advantage of enterokinase is that when the recognition sequence is fused to the C-terminus of the protein tag, it can be digested without amino acid residue.DDDK enterokinase is a tool enzyme. Enterokinase specifically cleaves peptides or proteins containing recognition sequences and is commonly used in biotechnology to perform specific cleavages of fusion proteins to release the protein of interest. DDDK enterokinase is highly effective and specific. In protein structure and function studies, it is used to accurately cleave the target protein from the fusion protein for subsequent analysis and experimentation. In the biopharmaceutical process, it can be used to prepare protein drugs with specific activities. The enterokinase recognition site on the plasmid serves as a recognition site, allowing the enterokinase to act on the induced proteins.[1,2]

Source

In addition, the low-cost and high-efficiency one-step protein separation method of histidine (His) labeling at the end of the protein and affinity purification of Nickel metal chelating column is also widely used in the separation and purification of rEK, with a yield of more than 50%. For the His-tag site, Choi et al. found that the enzyme activity of rECL was not changed after the C-terminus was His-labeled, but the enzyme activity was lost after the N-terminus was labeled by the Nickel metal chelating column.[3]

References

[1] ZHANG Xianghui,TAN Shuhua,LI Taiming. Research progress on the characteristics of enterokinase and its genetic engineering. CNKI, 2005

[2] Xu Liang, Wang Wenjing, Zu Xiangyang, Wang Xiaochun. Characteristics of enterokinase digestion of fusion protein Trx-Exendin-4. 《CNKI; WanFang", 2012

[3] Cao Zhifei, Li Xinping, Fan Kai, Li Jing, et al. Expression and purification of recombinant bovine enterokinase light chain gene in Pichia pastoris. 《CNKI; WanFang》，2006