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[Expression of recombinant ribosome inactivating protein MAP30 in E.coli and its biological activity]

Li-li Zhang1, Qian Ding, Jin-biao Zhan

  • 1Department of Biochemistry and Genetics, College of Medicine, Zhejiang University, Hangzhou 310058, China.

Zhejiang Da Xue Xue Bao. Yi Xue Ban = Journal of Zhejiang University. Medical Sciences
|June 15, 2010
PubMed
Abstract

Insights

Bitter melon

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Pharmacology

Background:

  • Momordica charantia L. (bitter melon) seeds contain ribosome inactivating proteins (RIPs).
  • MAP30 is a RIP with potential therapeutic applications.
  • Efficient production of bioactive MAP30 is crucial for further research.

Purpose of the Study:

  • To clone and produce recombinant ribosome inactivating protein MAP30 from bitter melon seeds.
  • To assess the biological activity and cytotoxicity of the recombinant MAP30 protein.

Main Methods:

  • Polymerase chain reaction (PCR) and T-A cloning were used to isolate the MAP30 gene.
  • The MAP30 gene was inserted into the pET30a vector for expression in E. coli.
  • Recombinant MAP30 expression was induced using IPTG, and protein activity was evaluated via MTT assay.

Main Results:

  • The cloned MAP30 sequences (nucleotide and amino acid) matched reported sequences.
  • SDS-PAGE confirmed the production of soluble recombinant MAP30.
  • The recombinant MAP30 exhibited higher cytotoxicity towards cancer cells compared to normal cells.

Conclusions:

  • The gene encoding MAP30 was successfully cloned and expressed in E. coli.
  • The recombinant MAP30 protein demonstrated significant bioactivity and selective cytotoxicity.

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