Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

[Human angiogenin: expression, purification, biological assay].

H Yang1, Y Q Zhang, Z Yan

  • 1Fourth Military Medical University Biotechnology Center, Xi'an, China. bio707@fmmu.edu.cn

Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology
|May 2, 2001
PubMed
Summary

Recombinant human Angiogenin (rhANG) was successfully expressed and purified. This protein can induce new blood vessel formation and degrade tRNA, showing its biological activity.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Antagonistic actions of Arabidopsis cryptochromes and phytochrome B in the regulation of floral induction.

Development (Cambridge, England)·1999
Same author

Intracisternal TRH analog increases gastrin release and corpus histidine decarboxylase activity in rats.

The American journal of physiology·1999
Same author

Co-encapsulation of Sertoli enriched testicular cell fractions further prolongs fish-to-mouse islet xenograft survival.

Transplantation·1999
Same author

Liposome-mediated immunotherapy against respiratory influenza virus infection using double-stranded RNA poly ICLC.

Vaccine·1999
Same author

Localization of a human nucleoporin 155 gene (NUP155) to the 5p13 region and cloning of its cDNA.

Genomics·1999
Same author

Effects of starvation and bowel resection on paracellular permeability in rat small-bowel mucosa in vitro.

Scandinavian journal of gastroenterology·1999

Area of Science:

  • Molecular Biology
  • Protein Expression
  • Biochemistry

Context:

  • Angiogenin (ANG) is a key protein involved in angiogenesis.
  • Efficient expression and purification of recombinant proteins are crucial for functional studies.
  • The His6-tag facilitates purification via affinity chromatography.

Purpose:

  • To clone and express recombinant human Angiogenin (rhANG) fused with a His6-tag.
  • To purify the expressed rhANG using Ni2(+)-NTA chelating resin.
  • To validate the biological activity of the purified rhANG.

Summary:

  • Angiogenin cDNA was reverse-transcribed and polymerase chain-reacted (RT-PCR), then cloned into the pRSETB vector for fusion expression.
  • The recombinant Angiogenin protein, with an N-terminal His6-tag, was expressed as an inclusion body at approximately 10% of total bacterial protein.

Related Experiment Videos

  • Following dissolution in 8 mol/L urea, the protein was purified using Ni2(+)-NTA chelating resin, leveraging the His6-tag's affinity for nickel ions.
  • Biological assays confirmed that the purified rhANG induced new blood vessel formation in the chorioallantoic membrane (CAM) and degraded transfer RNA (tRNA) in vitro.

    • Impact:

    • Provides a method for producing biologically active recombinant human Angiogenin.
    • Demonstrates the functional capabilities of purified rhANG in angiogenesis and tRNA degradation.
    • Contributes to the understanding of Angiogenin's role in biological processes.