Cloning of human HNA-3a gene and its expression in HEK-293 cells

ObjectiveTo construct an eukaryotic expression vector containing full-length sequences of human HNA-3a gene, explore an experimental method for detecting human anti-HNA-3a antibody, and observe the protein expression of HNA-3a.

Methods The primers were designed and synthesized according to human HNA-3a gene sequences in Genebank.The full-length of HNA-3a gene was reversely transcribed using RT-PCR, and cloned into pEGFP-N3 vector.The pEGFP-N3 expression vector containing human HNA-3a gene was transfected into the HEK-293 cells for transient expression using Lipofectamine^TM 2000 reagent, and the expression of the target gene was identified by Western blotting.

Results The eukaryotic expression vector containing the full-length sequences of human HNA-3a gene was successfully constructed, and the target gene was highly expressed in HEK-293 cells.

Conclusions The eukaryotic expression vector with high expression of HNA-3a protein is successfully constructed, which lays the foundation for the detection of the anti-HNA-3a antibody in human plasma.