›› 2019, Vol. 46 ›› Issue (1): 200-205.doi: 10.16431/j.cnki.1671-7236.2019.01.023

• Genetics and Breeding • Previous Articles     Next Articles

Study on Regulation of Gene Expression of Smad3 Gene by miR-23a in Pig

QIU Meiyu, ZHAI Tengjiao, HUANG Tao, LI Tao, SUN Jingli, SUN Xiaomei   

  1. College of Animal Science and Technology, Shihezi University, Shihezi 832000, China
  • Received:2018-06-01 Online:2019-01-20 Published:2019-01-19

Abstract:

To determine whether the miR-23a regulate Smad3 gene,wild-type and mutant-type dual-luciferase reporter vectors containing Smad3-3'-UTR (psiCHECKTM-2-W-Smad3-3'-UTR and psiCHECKTM-2-M-Smad3-3'-UTR) were constructed using Not Ⅰ and Xho Ⅰ,miR-23a mimics,miR-23a inhibitor and their negative control were transfected with or without dual luciferase reporter vectors in PK-15 cells,luciferase activity were assayed,the mRNA and protein expression levels of Smad3 gene were determined by Real-time PCR and Western blotting,respectively.The results showed that the luciferase activity in PK-15 cells transfected with wild-type dual-luciferase reporter vectors and miR-23a mimic was significantly lower than negative control (P < 0.05).In PK-15 cells transfected with miR-23a mimic,the expression levels of Smad3 mRNA and Smad3 protein were significantly down-regulated (P < 0.05).While there was no significant difference in the expression level of Smad3 protein between PK-15 cells tansfected with miR-23a inhibitor and its negative control (P > 0.05).The results indicated that miR-23a could regulate directely the expression of its target gene Smad3.

Key words: miR-23a; Smad3 gene; expression regulation

CLC Number: