Preparation of hMan2c1 transgenic mice

Objective To develop hMan2c1 transgenic mice for studying the biological significance of MAN2C1 in vivo. Methods Recombinant expression plasmid pIRES2-EGFP-hMan2c1 was constructed. The expression of pIRES2-EGFP-hMan2c1 was indicated in COS-7 cells and then injected into the zygotes of ICR mice to prepare transgenic mice. Genomic PCR was performed to analyze integration of the target gene into the genomic DNA of the host. RT-PCR and Western blot were performed to detect expression of hMan2c1 in the transgenic mice. Results Out of the 116 first generation mice, 7 were genomic PCR positive for hMan2c1. Out of the 20 F1 mice examined, 9 were genomic PCR positive for hMan2c1. Out of the 21 F2 mice detected, 16 were genomic PCR positive for hMan2c1. RT-PCR and Western blot for hMan2c1 expression in mouse tail tissue showed that 4 were positive in the 7 transgenic mouse lines. Conclusion Four hMan2c1 transgenic mouse lines have been developed, which can be used for studying the biological significance of MAN2C1 in vivo.