OBJECTIVE The aim of this research was to clone and express the antigen of the previously prepared monoclonal antibody named M4G3. METHODS Western blots were used to screen a breast cancer cell line that overexpresses...OBJECTIVE The aim of this research was to clone and express the antigen of the previously prepared monoclonal antibody named M4G3. METHODS Western blots were used to screen a breast cancer cell line that overexpresses the M4G3-associated antigen. A λ, zap cDNA expression library of breast cancer cells was constructed and screened using M4G3 as a probe to clone the antigen. The positive clones were subcloned and identified by homologous comparison using BLAST. RESULTS The λ zap cDNA expression library had 1.0×10^6 independent clones. Fifteen positive clones were isolated following 3 rounds of immunoscreening and identified as being from Mycoplasma pulmonis. CONCLUSION The specific antigen that matched the monoclonal M4G3 antibody is an unknown protein of M. pulmonis. This work is helpful for the further study of the association of M. pulmonis infection with breast cancer.展开更多
基金This work was supported by the National Natural Science Foundation of China （No.30370553）.
文摘OBJECTIVE The aim of this research was to clone and express the antigen of the previously prepared monoclonal antibody named M4G3. METHODS Western blots were used to screen a breast cancer cell line that overexpresses the M4G3-associated antigen. A λ, zap cDNA expression library of breast cancer cells was constructed and screened using M4G3 as a probe to clone the antigen. The positive clones were subcloned and identified by homologous comparison using BLAST. RESULTS The λ zap cDNA expression library had 1.0×10^6 independent clones. Fifteen positive clones were isolated following 3 rounds of immunoscreening and identified as being from Mycoplasma pulmonis. CONCLUSION The specific antigen that matched the monoclonal M4G3 antibody is an unknown protein of M. pulmonis. This work is helpful for the further study of the association of M. pulmonis infection with breast cancer.
