[Cloning and expression of human Runx3 gene obtained from human peripheral blood CD8(+) T lymphocyte].
OBJECTIVE
Runx3, a type of Runt family member, plays an important role in immune regulation. In this study, we cloned and analyzed the cDNA encoding human Runx3 from T lymphocyte, expressed Runx3 protein in E.coli system, and studied the relation between Runx3 and some immune disorders or tumors.


METHODS
The CD8(+) T were isolated from human peripheral blood with MACS, Runx3 cDNA was amplified by RT-PCR and cloned into pMD19-T vector, and recombinant was transformed into competent cells DH5alpha and recombinant sequencing were performed. The identical was subcloned into pQE30 vector and expressed in E.coli M15. The fusion protein was identified by Western blot.


RESULTS
The 1,248 bp fragment amplified by RT-PCR was the same as the anticipated one in size and encodes 415 amino acids. Runx3 protein was gained.


CONCLUSIONS
Human Runx3 gene was cloned and expressed in E.coli system successfully, which brought a foundation for further research on its biological function.
Li
Ya-zhen
YZ
Department of Immunology, Institute of Clinical Laboratory Medicine, Jiangsu University, Zhenjiang, China. lyz-0115@163.com
Wang
Sheng-jun
SJ
Shi
Yan
Y
Dai
Xiao-li
XL
Peng
Su-fang
SF
Su
Zhao-liang
ZL
Shao
Qi-xiang
QX
Xu
Hua-xi
HX
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D050677:Core Binding Factor Alpha 3 Subunit / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment
D050677:Core Binding Factor Alpha 3 Subunit; C493729:Runx3 protein, human