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Updated: May 11, 2026
Production of Monoclonal Antibodies Targeting Aminopeptidase N in the Porcine Intestinal Mucosal Epithelium
Published on: May 18, 2021
[Preparation of monoclonal antibody against phosphinothricin acetyltransferase]
Xudong Gao1, Yongzhi Wang, Shengfeng Shi
1College of Horticulture, Northeast Agricultural University, Harbin 150030, China. eo_777654@sina.com
Objective:
To express phosphinothricin acetyltransferase (PAT) with biological activity and prepare monoclonal antibodies against PAT.
Methods:
The full length bar gene was cloned by PCR and inserted into prokaryotic expression vector pET28a⁺. The recombinant plasmid pET28-bar was transformed into E.coli BL21(DE3), and under the induction of IPTG, PAT was expressed. The expressed protein was purified by Ni⁺; affinity chromatography to analyze its activity. The purified PAT was used to immunize BALB/c mice, and then the spleen cells from the immunized mice were fused with Sp2/0 cells. The hybridoma clones secreting antibodies against PAT were isolated by indirect ELISA and then subcloned.
Results:
Soluble PAT was expressed in E.coli. The purified PAT had the activity of acetyltransferase. We totally prepared 9 hybridoma cell lines which secreted specific anti-PAT monoclonal antibodies.
Conclusion:
The expressed recombinant PAT can be used for biological reagent to prevent and relieve herbicide damage. Monoclonal antibodies against PAT may be used to detect the transgenic products.
