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- Title
Construction and effect test of the eukaryotic expression vector pcDNA3.1(+)-MyoD-HA-His.
- Authors
DU Chao; YU Leiye; SUN Guojie; WANG Yingze
- Abstract
In order to conveniently detect and purify the myogenic factor MyoD exo-expressed in the non-muscle cells, and to compare with the expression level of other myogenic factors cotransfected with MyoD, the coding sequence (CDS) of MyoD was cloned and inserted into pcDNA3. 1(+)-HA-His, an eukarytic expression vector. Sequencing result confirms that the insert is correct and forms the same ORF with the HA and its sequences. Furthermore, Western blot shows that the fusion protein MyoD-HA expresses with a high level for the addition of kozak sequence GCCGCCACC, and myogenic conversion demonstrates the fusion protein MyoD-HA can activate the target genes effectively.
- Subjects
MYOBLASTS; CHIMERIC proteins; NUCLEOTIDE sequence; GENE targeting; EUKARYOTES; CYTOGENETICS; CELLS
- Publication
Journal of Hebei University of Science & Technology, 2014, Vol 35, Issue 2, p149
- ISSN
1008-1542
- Publication type
Article
- DOI
10.7535/hbkd.2014yx02007