Welcome to Chinese Agricultural Science Bulletin,

Chinese Agricultural Science Bulletin ›› 2009, Vol. 25 ›› Issue (3): 13-17.

Special Issue: 生物技术 畜牧兽医

• 畜牧兽医科学 • Previous Articles     Next Articles

Construction and identification of GFP reporter vectors driven by chicken CVH gene promoters

  

  • Received:2008-10-13 Revised:2008-12-01 Online:2009-02-05 Published:2009-02-05

Abstract: In chicken, the distribution of CVH transcripts is restricted to the germ cell lineage, making it a useful indicator of PGCs. In order to label live PGCs , we cloned and characterized the chicken1.6kb CVH gene promoter regions. TATA box and CAAT box-like elements were found in the sequence,and GC domains were rich in the far upsteam of the promoter. The GFP reporter vectors (pCVH-EGFP) driven by CVH gene promoters was constructed. The blastodermal cell and CEF cell were transfected respectively by LipofectamineTM2000 with pCVH-EGFP for transient expression. The GFP-labeled positive blastodermal cells were first observed 12 h post-tranfection and increased with the post-transfection time (in 24 h). GFP-labeled and non-labeled cells from X blastoderm were large and round with the same characters of morphology. The GFP protein was not expressed in CEF cell .Our results suggested that the CVH promoter regions have specific activity in stage X blastoderm. It is possiblely to develop a method for labeling chicken PGCs with GFP and subsequently to sort viable GFP-positive PGC from somatic cell by flow cytometry. This should be extremely useful for applications of PGC transplantation