[Construction of eukaryotic expression vector of LYRM1, a novel gene related with human obesity, and establishment of its stable transfected 3T3-L1 cell line].
To construct the eukaryotic expression vector of LYRM1 and to transfect it to preadipocytes cell line 3T3-L1.



The 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.



The 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.



The eukaryotic expression vector of LYRM1 has been successfully established and the stably transfected 3T3-L1 cell line also established.
Qiu
Jie
J
Department of Newborn Infants, Nanjing Children's Hospital of Nanjing Medical University, Nanjing 210008, China.
Zhang
Min
M
Zhou
Xiao-Yu
XY
Cheng
Rui
R
Cao
Xing-Guo
XG
Wang
Bin
B
Guo
Xi-Rong
XR
D041721:3T3-L1 Cells; D000273:Adipose Tissue / Q000378:metabolism; D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000096:biosynthesis* / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D009765:Obesity / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D051017:Apoptosis Regulatory Proteins; C538836:LYRM1 protein, human; D011994:Recombinant Proteins