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[The cloning and expression of soluble HLA-B27 gene]
Objective:
To clone and express the soluble HLA-B27 (sB27) gene in human B lymphoid cell line.
Methods:
Soluble B27 cDNA was constructed by 3 consecutive rounds of PCR and then cloned into the RSV5neo vector. The construct was transfected into C1R cells.
Results:
Restriction enzyme digestion and DNA sequencing confirmed that the construct was the HLA-B27 gene lacking exon 5 which encodes the transmembrane domain of HLA molecule. The expression of aHLA-B27 was verified successfully in the culture supernatant of C1R cells. The level of sHLA-B27 changed with variable culture conditions, such as temperature and concentration of fetal calf serum.
Conclusion:
The sHLA-B27 molecules can be generated by alternative splicing of HLA-B27 pre-mRNA. Our construct provide a useful model for studying the effect of environmental factors such as bacterial infections and cytokine stimulation on the expression of soluble HLA-B27 molecule. It will also provide clues for the role of HLA-B27 molecule in the pathogenesis of spondyloarthropathies.