Abstract: The aim of this experiment was to construct overexpression vectors of BMP6 and BMPR1B genes in Aohan fine-wool sheep, and then introduce them into fibroblast cells, to analyze the changes of mRNA and protein expression of the two genes after transfection and the effect of gene interaction on hair follicle. The 40-day-old embryo of Aohan banner fine-wool sheep was collected, RNA was extracted and reverse transcribed into cDNA. The purified target fragment was linked to PCDNA3.1 by SoSo Kit after PCR reaction. After restriction endonuclease identification, the bacteria was transferred to E. coli DH5α natural competence, shaken and picked to expand the single colony culture and vibration culture and pick to expand the single colony culture. The plasmids pcDNA3.1-BMP6 and pcDNA3.1-BMPR1B were co-transfected into fibroblasts. After transfection, the expression level was measured and the relationship between the two genes were explored. The results showed that compared with the single transfected group, the expression of BMPR1B gene and BMPR1B protein was significantly increased in co-transfected fibroblasts(P<0.01),but the expression of BMP6 gene and BMP6 protein in co-transfection group had no significant difference(P>0.05). Therefore, BMP6 gene promotes the expression of BMPR1B gene, and BMPR1B gene has little effect on the expression of BMP6 gene.

Key words: Aohan fine-wool sheep, Homologous recombination technology, Fibroblasts, Real-time PCR, Western Blot

CLC Number: 

• Q78