[Cloning and expression of human IL-1R II gene].
OBJECTIVE
To clone human IL-1R II cDNA and construct its recombinant retrovirus vector so as to explore its role in IL-1R II related diseases.


METHODS
Human IL-1R II cDNA was amplified by RT-PCR from peripheral blood mononuclear cells (PBMCs) and inserted into the vector PET22b to construct recombinant vector PET22b-IL-R II. The recombinant was transfected into E. coli BL21 and expressed under IPTG induction. Expressed products were detected by Western blot. In addition, human IL-1R-II cDNA was subcloned into retrovirus vector LZRSPBMN and transfected into 293 cells by calcium phosphate precipitation. IL-1R II expression was detected by immunohistochemical staining.


RESULTS
IL-1R II cDNA with 1,203 bp was amplified by RT-PCR from human PBMCs. The recombinant of this cDNA could be expressed in E. coli,which was confirmed by Western blot results. Immunohistochemistry detection showed IL-1R II protein was expressed in 293 cells.


CONCLUSIONS
Human IL-1R II gene was cloned successfully. PET22b-IL-1R II and LZR-IL-1R II were constructed and the recombinant protein IL-1R II was expressed in E.coli BL21. The results reported herein lay the foundation for further research on the role of IL-1R II in certain diseases.
Hu
Yan-qiu
YQ
Department of Obstetrics and Gynecology, the First Affiliated Hospital, Nanjing 210029, China. huyanqiu78@163.com
Liu
Jia-yin
JY
Lu
Chun
C
Wang
Shu-yu
SY
D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D007668:Kidney / Q000166:cytology; D010957:Plasmids; D017472:Receptors, Interleukin-1 / Q000096:biosynthesis* / Q000235:genetics; D053574:Receptors, Interleukin-1 Type II; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012190:Retroviridae / Q000235:genetics*; D014162:Transfection
D018076:DNA, Complementary; D017472:Receptors, Interleukin-1; D053574:Receptors, Interleukin-1 Type II; D011994:Recombinant Proteins