Abstract: The study aimed to identify the expression of Hoxa5 and BMPR1B gene plasmid and the interaction between the two genes was studied by means of expression changes. Firstly, a pair of primers were desigened by RNA extraction and referring to Hoxa5 and BMPR1B gene sequence information of Aohan fine wool sheep in GenBank, and the Hoxa5 and BMPR1B gene fragment were amplified by PCR method. The obtained Hoxa5 and BMPR1B gene were ligated into pEASYTM-T1 vector to construct pEASYTM-T1-Hoxa5 and pEASYTM-T1-BMPR1B recombinant plasmid and transformed into E.coli DH5α competent cell. The plasind was identified by restriction enzyme digedtion. The recombinant plasimd pcDNA3.1-Hoxa5 and pcDNA3.1-BMPR1B was constructed and transformed into E.coli DH5α competent cell. Then pcDNA3.1-Hoxa5 and pcDNA3.1-BMPR1B plasmid were cotransfection into fibroblasts, and qRT-PCR was used to detect the expression level of its gene. As the results showed that, pcDNA3.1-Hoxa5 and pcDNA3.1-BMPR1B plasmid cotransfection successfully and identified by enzyme and sequencing. The expression level of BMPR1B gene in fibroblasts was lower than in the control group and expression level of Hoxa5 gene in fibroblasts was higher than in the control group. The plasmid was constructed and cotransfection into fibroblasts successfully. The expression level of Hoxa5 gene increased significantly, and the expression of BMPR1B gene decreased. Therefore, Hoxa5 gene inhibited the expression of BMPR1B gene, and BMPR1B gene promoted the expression of Hoxa5 gene.These results would lay a foundation for the further research.

Key words: Aohan fine wool sheep, Fibroblats, Hoxa5, BMPR1B, Gene interaction, Plasmid construction, RT-PCR, Western Blot

CLC Number: 

• Q78

• S826