[Construction of mic2/CD99 gene vector and its transfection in Hodgkin lymphoma L428 cell line].
OBJECTIVE
To construct a eukaryotic expression vector of CD99 gene for transfection into Hodgkin lymphoma L428 cells.


METHODS
The full-length cDNA of CD99 gene was amplified from Jurkat cells by RT-PCR and cloned into the pcDNA3.1(+) vector and transfected into L428 cell line using Lipofextamine 2000. The sequence of CD99 mRNA in the transfected cells was confirmed by restriction endonuclease digestion and DNA sequencing, and the expression of CD99 protein was identified using immunocytochemistry.


RESULTS
A gene fragment of 558 bp was amplified from the transfected cells and the sequence was verified by DNA sequencing. Immunocytochemistry identified the presence of CD99 expression in the transfected cells.


CONCLUSIONS
A eukaryotic expression vector pcDNA3.1(+)-CD99 is successfully constructed and stably expressed in L428 cell line.
Huang
Zuo-ping
ZP
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. zuoping68@126.com
He
Ying
Y
Zhou
Xin-hua
XH
Han
Xi-qing
XQ
Wu
Zi-Qun
ZQ
Zhang
Yan
Y
Wen
Zong-Hua
ZH
Zhao
Tong
T
D000071443:12E7 Antigen; D015703:Antigens, CD / Q000096:biosynthesis* / Q000235:genetics; D001483:Base Sequence; D015815:Cell Adhesion Molecules / Q000096:biosynthesis* / Q000235:genetics; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D005822:Genetic Vectors / Q000235:genetics*; D006689:Hodgkin Disease / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D019169:Jurkat Cells; D008969:Molecular Sequence Data; D014162:Transfection
D000071443:12E7 Antigen; D015703:Antigens, CD; C061828:CD99 protein, human; D015815:Cell Adhesion Molecules