Gene targeting combined with culture of chicken embryonic stem cells is one of the best strategies to produce chicken oviduct bioreactor. To study the construction of a vector with Cre recombinase recognition site loxp mutant for chicken Ovalbumin gene targeting in embryonic stem cells, the 1.7 kb fragment including chicken partly gene 5′flanking sequence, and exon 1, intron1 and partly exon 2 and the 6.1 kb fragement including OV gene from exon 2 to exon 8 were cloned. Using two chicken OV gene DNA fragments, the internal ribosome entry site (IRES) of the (ECMV) virus and synthesized oligonucleotides, pSSC-m2/71EGFP66IRES-OV7.8 was designed and constructed as a gene targeting vector. Then the shuttle vector pMD-m2/66-MCS-IFN-MCS-Loxp71 was constructed. Structures of the two constructs were identified by restrictive digestion analysis and partial sequencing. To check the function of two Cre“exchange cassette” , the targeting construct and the lineal shutter construct were tested in E.coli BM25.8 that express the Cre recombinase constitutively. The results indicated that this targeting construct system can be used in the study of chicken oviduct bioreactor[Acta Zoologica Sinica 51(4): 685–690, 2005].
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