ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2005, Vol. 36 ›› Issue (10): 987-990.doi:

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Molecular Cloning and Sequence Analysis of Putative U3-R Sequence of cENS-2 Gene and Construction of Expression Vector

ZHANG Chuan-sheng;GENG Li-ying; DU Li-xin   

  1. 1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100094,China；2Department of Animal Socience,Hebei Normal University of Science and Technology,Changli 066600,China;3.Department of Life Science,Hebei Normal University of Science and Technology,Changli 066600,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2005-10-25 Published:2005-10-25

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Abstract: This report describes construction of specific labeling vector in CES cell. The putativeU3-R sequence regions of 1.2 kb of chicken Embryonic Normal Stem cell gene 2 was amplified by PCR and successfully sequenced. Based on sequence analysis and alignments of nucleotides sequence, found many factor or protein binding sites such as AP-1 ,GATA-1and an important elements: B region which exhibits a strong enhancer activity in CES and differentiated cells. The CMV promoter was cut off from pEGFP-N1, the putative U3-R fragment was inserted just upstream of GFP . Results of restriction digestion and DNA sequencing showed that the vector for expressing in CES cell was successfully constructed.