Abnormal expression of developmentally important genes leads to inadequate reprogramming development of NT embryo which subsequently causes failure in animal cloning. To identify reprogramming-associated genes in rabbit NT embryo, mRNA differential display has been developed and SPEDDRT-PCR method was set up successfully to overcome the paucity of the biological materials. Using this modified technique, the mRNA content of rabbit NT embryos at different developmental stages (from oocyte to 8～16-cell embryo) were compared, eighty differential displayed bands at different stage in rabbit NT embryo were isolated. A028 amplicon was confirmed by reverse Northern blot. Nucleotide sequences were determined for these cDNAs and database searches identified using NCBI BLAST program. The data suggested that A028 displayed high homology (93%) to CstF3 gene for cleavage stimulation factor, which involved in pre-mRNA 3′-end processing and is required for progression through mitosis. This gene was probably related with preimplantation embryo development, and might play important roles in development of rabbit NT embryo. Of seven organ tissues examined by Northern blot analysis, A028 was only found in ovary. This work paves the way of cloning of the full length of A028 cDNA and further study on gene function.