Construction of mic2/CD99 gene vector and its transfection in Hodgkin lymphoma L428 cell line.
- Author:
Zuo-ping HUANG
1
;
Ying HE
;
Xin-hua ZHOU
;
Xi-qing HAN
;
Zi-Qun WU
;
Yan ZHANG
;
Zong-Hua WEN
;
Tong ZHAO
Author Information
- Publication Type:Journal Article
- MeSH: 12E7 Antigen; Antigens, CD; biosynthesis; genetics; Base Sequence; Cell Adhesion Molecules; biosynthesis; genetics; Cell Line, Tumor; Cloning, Molecular; Genetic Vectors; genetics; Hodgkin Disease; metabolism; pathology; Humans; Jurkat Cells; Molecular Sequence Data; Transfection
- From: Journal of Southern Medical University 2009;29(12):2407-2409
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct a eukaryotic expression vector of CD99 gene for transfection into Hodgkin lymphoma L428 cells.
METHODSThe 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.
RESULTSA 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.
CONCLUSIONA eukaryotic expression vector pcDNA3.1(+)-CD99 is successfully constructed and stably expressed in L428 cell line.
