[Construction of eukaryotic expression vector for HPC2 and its expression in HEK293 cells].
OBJECTIVE
To construct the eukaryotic expression vector for HPC2 for expression in HEK293 cells.


METHODS
HPC2 from pcDNA3/HPC2 were inserted into the flag-tagged vector pcDNA3-flag by subcloning method. The recombinant plasmid pcDNA3-flag/HPC2 was then transfected into HEK293 cells using a routine lipofectamine method. The cell lysate was used for Western blotting to examine the expression of the target protein.


CONCLUSIONS
Double restriction enzyme digestion and DNA sequencing indicated successful construction of the eukaryotic expression vector for HPC2 and the fusion protein was highly expressed in HEK293 cells, which provides an important basis for functional study of HPC2.
Tan
Kang-lian
KL
Key Laboratory of Functional Proteomics of Guangdong Province, Southern Medical University, Guangzhou 510515, China. collinton@126.com
Li
Zhi-jie
ZJ
Liu
Jing-hua
JH
Huang
Hao
H
Tang
Jing
J
Deng
Peng
P
Jiang
Yong
Y
D002460:Cell Line; D003001:Cloning, Molecular; D005057:Eukaryotic Cells / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D008025:Ligases; D010957:Plasmids / Q000235:genetics; D063146:Polycomb-Group Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics; D044767:Ubiquitin-Protein Ligases
D063146:Polycomb-Group Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D044767:Ubiquitin-Protein Ligases; D008025:Ligases; C570427:CBX4 protein, human