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Updated: Aug 14, 2026
Generation of Murine Monoclonal Antibodies by Hybridoma Technology
Published on: January 2, 2017
Production and characterization of a monoclonal antibody to the v-mos oncogene protein
1Division of Cell Biology, Fujita Health University, Aichi, Japan.
Abstract:
Valuable information about proto-oncogenes and their physiological functions has been obtained by studying their expression in normal cells. However, the protein product of the c-mos gene, the cellular homologue of the transforming gene (v-mos) of Moloney murine sarcoma virus, has not been detected in normal mouse cells or tissues. Here, we have constructed a v-mos expression vector, pRI-delta mos, which directs the synthesis of a truncated v-mos gene product, a protein A fusion protein. Using the truncated v-mos oncoprotein produced in Escherichia coli as immunogen, we prepared anti-v-mos monoclonal antibodies (MAbs). In immunoblotting assays, the MAb was reactive with v-mos oncoprotein and detected bands at 43 KDa or 39 kDa in the tissue extract of mouse testes or ovaries, respectively, in which the c-mos protooncogene mRNA is expressed. These results demonstrate that the v-mos MAb obtained is suitable for elucidating the physiological functions of v-mos gene product and may also be utilized to detect c-mos gene product at the cellular level.
Insights
Researchers developed monoclonal antibodies (MAbs) to detect the c-mos proto-oncogene product in normal mouse cells. These MAbs successfully identified the c-mos protein in mouse testes and ovaries, revealing its physiological functions.
Area of Science:
- Molecular Biology
- Oncology
- Immunology
Background:
- Proto-oncogenes provide valuable information on normal cellular functions.
- The c-mos gene product, a homologue of the Moloney murine sarcoma virus transforming gene (v-mos), has not been detected in normal mouse cells.
- Understanding c-mos function is crucial for comprehending cellular processes and oncogenesis.
Purpose of the Study:
- To develop tools for detecting the c-mos gene product in normal mouse tissues.
- To investigate the physiological roles of the c-mos proto-oncogene.
- To generate specific antibodies against the v-mos oncoprotein for detection purposes.
Main Methods:
- Construction of a v-mos expression vector (pRI-delta mos) for synthesizing a truncated v-mos gene product fused to protein A.
- Production of a truncated v-mos oncoprotein in Escherichia coli.
- Generation of anti-v-mos monoclonal antibodies (MAbs) using the purified oncoprotein as an immunogen.
- Immunoblotting assays to test MAb reactivity with the v-mos oncoprotein and detect c-mos in mouse tissue extracts.
Main Results:
- The generated MAb specifically reacted with the v-mos oncoprotein.
- The MAb detected protein bands of 43 kDa in mouse testes and 39 kDa in mouse ovaries.
- These tissues were known to express c-mos proto-oncogene mRNA, confirming the detection of the c-mos gene product.
- The results indicate the successful identification of the c-mos protein at the cellular level.
Conclusions:
- The developed anti-v-mos MAb is a valuable tool for detecting the c-mos gene product in normal mouse cells.
- This MAb can be used to elucidate the physiological functions of the c-mos proto-oncogene.
- The study provides a method for cellular-level detection of c-mos, advancing research in proto-oncogene function and potential oncogenic pathways.
