Format

Send to

Choose Destination
See comment in PubMed Commons below
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2014 Jun;22(3):807-11. doi: 10.7534/j.issn.1009-2137.2014.03.043.

[Construction and expression of fusion protein TRX-hJagged1 in E.coli BL21].

[Article in Chinese]

Author information

  • 1Department of Hematology, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi.
  • 2Department of Traditional Chinese Medicine, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi.
  • 3Department of Hematology, Tangdu Hospital of The Fourth Military Medicol University, Xian 710038, Shaanxi. E-mail: liangym@fmmu.edu.cn.

Abstract

This study was purposed to construct prokaryotic expression vector and to investigate the expression of Notch ligand Jagged1 in E.coli. An expression vector pET-hJagged1 was constructed, which can be inserted in Jagged1 with different lengths, but the DSL domain of human Jagged1 should be contained. Then the recombinant plasmids were transformed into the competent cell of E.coli BL21, and the expression of the fusion protein was induced by IPTG. Fusion protein was purified from the supernatant of cell lysates via the Nickel affinity chromatography. The results showed that prokaryotic expression vectors pET-hJagged1 (Bgl II), pET-hJagged1 (Hind I) and pET-hJagged1 (Stu I) were successfully constructed, but only pET-hJagged1 (Stu I) could express the soluble TRX-hJagged1. The purified TRX-Jagged1 protein could be obtained via the Nickel affinity chromatography, and then confirmed by Western Blot. It is concluded that prokaryotic expression vector pET-hJagged1 is successfully constructed, but only pET-hJagged1 (Stu I) can express the soluble TRX-hJagged1 and the TRX-Jagged1 fusion protein is obtained through the prokaryotic expression system, which laid a solid foundation for further to explore the effects of Jagged1 in hematopoietic and lymphoid system.

PMID:
24989299
[PubMed - indexed for MEDLINE]
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Loading ...
    Support Center