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Updated: Sep 30, 2026
Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
Published on: February 25, 2010
Expression of Smad4 Gene in Pichia pastoris and Identification of the Protein
Yi-Ping Liu1, Xiao Yang, Xiao-Ling Xu
1Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100071, China. huangc@nic.bmi.ac.cn
Abstract:
Smad4 is a novel tumor suppressor gene which is mutated or deleted in about 50% of pancreatic carcinoma and 30% of colon cancer. SMAD4 was expressed in Pichia pastoris and was characterized. The molecular weight of the expression product was shown to be about 67 kD, and 13 amino acids in N terminal were determined and were identical with the putative sequence from Smad4 cDNA, and it was able to specifically react with the antibody against SMAD4.
Insights
Smad4, a tumor suppressor gene, is crucial in pancreatic and colon cancers. Researchers successfully expressed and characterized SMAD4 in Pichia pastoris, confirming its identity and reactivity.
Area of Science:
- Molecular Biology
- Oncology
- Biochemistry
Background:
- Smad4 is a critical tumor suppressor gene.
- Mutations or deletions in SMAD4 occur in approximately 50% of pancreatic carcinomas and 30% of colon cancers.
- Understanding SMAD4's function is vital for cancer research.
Purpose of the Study:
- To express and characterize the SMAD4 protein.
- To confirm the identity and properties of the expressed SMAD4 protein.
Main Methods:
- Gene expression in Pichia pastoris.
- Protein molecular weight determination.
- N-terminal amino acid sequencing.
- Immunological reactivity testing with SMAD4 antibody.
Main Results:
- SMAD4 was successfully expressed in Pichia pastoris.
- The expressed protein had a molecular weight of approximately 67 kD.
- The N-terminal 13 amino acids matched the predicted sequence from SMAD4 cDNA.
- The expressed protein specifically reacted with an antibody against SMAD4.
Conclusions:
- The successful expression and characterization of SMAD4 in Pichia pastoris provide a reliable source for further functional studies.
- The confirmed identity and reactivity of the expressed SMAD4 protein are essential for its use in cancer research and therapeutic development.
