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Cloning of Mouse SRY-Box Gene SOX1 and Constructing of Eukaryotic Expression Vector pEGFP-C3-SOX1

GUO Jian-xin~*,XU Yu-ming,ZHAO Guo-qiang,ZHANG Rui,SONG Bo,ZHANG Shi-jie,ZHANG Shi-feng,LI Jing-hong.~*Department of Neurology,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052,China  
Objective:To clone the entire coding sequence and construct a mouse sox1 expression vector PEGFP-C3-sox1 for studying the effect of SOX1 on neural differentiation of stem cells.Methods: The fragment of sox1 containing EcoRI and HindIII was amplified by RT-PCR from the mouse earlier embryo and then cloned into pGEM-T Easy vector.The recombinant plasmid was identified by restriction-endonuclease digestion and DNA sequencing.The fragment of sox1 was digested by EcoRI and HindIII and subclonded into the expression vector pEGFP-C3,which was denominated with pEGFP-C3-SOX1,and then the recombinant plasmid was identified by restriction-endonuclease double digestion.Results: Restriction-endonuclease digestion and DNA sequencing showed that the recombinant plasmid pEGFP-C3 contained the fragment of sox1.Conclusion:The entire coding sequence of mouse SRY-Box gene SOX1 was successfully cloned and sox1 expression vector pEGFP-C3-sox1 was constructed.
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