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Construction and confirmation of a recombinant eukaryotic expression plasmid pBABE-hygro-PLAP-1

Zhang, P.-P.; Li, S.; Yang, P.-S.; Sun, J.

Shanghai Kou Qiang Yi Xue 19(6): 635-640

2010

ISSN/ISBN: 1006-7248
PMID: 21431266
Document Number: 646057
To construct and confirm a recombinant eukaryotic expression plasmid pBABE-hygro-PLAP-1 containing periodontal ligament-associated protein(PLAP)-1 and a known FLAG sequence. Firstly, the targeted DNA fragment was amplified from pCAG_PLAP-1 by PCR, then PCR product was inserted into p3×FLAG-CMV-10 vector with a known sequence of FLAG. Secondly, plap-1 gene and FLAG were obtained by PCR amplification from p3×FLAG-PLAP-1, then PCR product was inserted to proper sites of vector pBABE-hygro. Finally, the recombinant plasmid pBABE-hygro-PLAP-1 was confirmed by agarose gel electrophoresis and DNA sequence analysis. The construction of recombinant eukaryotic expression plasmid pBABE-hygro-PLAP-1 was confirmed through restriction enzyme maping analysis and DNA sequencing. The recombinant plasmid of pBABE-hygro-PLAP-1 is successfully constructed. It can be applied to plap-1 gene function research in the future. Supported by Natural Science Foundation of Shandong Province (Grant No.2R2010HM050).

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