[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.


CONCLUSIONS
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.
Huang
F
F
Department of Rheumatology, PLA General Hospital, Beijing 100853, China.
Zhang
J
J
Yu
D
D
D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression; D015796:HLA-B27 Antigen / Q000096:biosynthesis* / Q000235:genetics*; D006801:Humans
D015796:HLA-B27 Antigen