Construction of eukaryotic expression vector of LYRM1, a novel gene related with human obesity, and establishment of its stable transfected 3T3-L1 cell line.
- Author:
Jie QIU
1
;
Min ZHANG
;
Xiao-Yu ZHOU
;
Rui CHENG
;
Xing-Guo CAO
;
Bin WANG
;
Xi-Rong GUO
Author Information
- Publication Type:Journal Article
- MeSH: 3T3-L1 Cells; Adipose Tissue; metabolism; Animals; Apoptosis Regulatory Proteins; biosynthesis; genetics; Genetic Vectors; Humans; Mice; Obesity; genetics; Recombinant Proteins; biosynthesis; genetics; Transfection
- From: Journal of Zhejiang University. Medical sciences 2009;38(5):493-497
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the eukaryotic expression vector of LYRM1 and to transfect it to preadipocytes cell line 3T3-L1.
METHODSThe complete coding sequence of LYRM1 gene was amplified by RT-PCR from human omental adipose tissue and was subcloned into eukaryotic expression vector pcDNA(TM)3.1/myc-His B. The recombinant plasmid was then transfected into 3T3-L1 preadipocytes by liposome. After screening culture by G418, stable transfected 3T3-L1 cell line was established. The expression of LYRM1 protein was identified by Western blot.
RESULTThe recombinant plasmid was confirmed by PCR, restriction enzyme digestion and sequencing. The stable transfected 3T3-L1 cell line was established and the LYRM1 protein was expressed successfully.
CONCLUSIONThe eukaryotic expression vector of LYRM1 has been successfully established and the stably transfected 3T3-L1 cell line also established.
