Abstract: 【Objective】 The genes of Oct4, Sox2, Klf4 and c-Myc were cloned, the pMD19-T-oct4, pMD19-T-sox2, pMD19-T-klf4, and pMD19-T-c-myc recombinant plasmids were constructed, and then the pcDNA3-Oct4, pcDNA3 -Sox2, pcDNA3-Klf4 and pcDNA3-c-Myc recombinant plasmids containing T7 promoter were constructed. According to the in vitro transcription, the four mRNA transcription factors were obtained, and it was made to express stably in Xuhuai goat fibroblasts. 【Method】Through the testicular, skin and small intestine tissue, the Oct4, Sox2, Klf4, and c-Myc genes coding sequence were obtained by RT-PCR, and were cloned into pMD19-T vector, and then cloned into the eukaryotic expression pCDNA3.0 vector that contains T7 promoter, and then the pcDNA3-Oct4, pcDNA3-Sox2, pcDNA3-Klf4 and pcDNA3-c-Myc recombinant plasmids were constructed. Recombinant plasmids were linearized using XhoI and XbaI, in vitro transcription of the transcription factors was made to obtain the mRNA using the instructions of in vitro transcription, the mRNA was detected, its stability and concentration were determined, Lipofectamine 2000 transfection was carried out according to the proportion of Lipofectamine﹕mRNA=1﹕1. Western blot technique and immunofluorescence assay were used to detect the mRNA expression in Xuhuai goat fibroblasts 24 h post-transfection. 【Result】 The length of Xuhuai goat Oct4, Sox2, Klf4 and c-Myc genes coding sequence were 1 083 bp, 962 bp, 1 434 bp and 1 320 bp, and confirmed by TA cloning, the sequences similarity of coding sequences of sheep, human, cattle and pig were above 89%. The mRNA of four transcription factors by transfection to Xuhuai goat fibroblasts were localized in the nucleus. The mRNA of four transcription factors expression in fibroblasts proteins was consisted with the expected size, 38 kd, 34 kd, 50 kd and 48 kd. 【Conclusion】The Xuhuai goat Oct4, Sox2, Klf4 and c-Myc transcription factors were cloned successfully, and the four mRNA can express in fibroblasts stably, thus a foundation for further study of the function of the genes Oct4, Sox2, Klf4, c-Myc and goat somatic cell reprogramming is established.

Key words: Xuhuai goat , transcription factors , in vitro transcription , mRNA