Inhibitory effect of RNA interference of MSTN gene expression on the downstream genes in Schizopygopsis pylzovi
Received:January 08, 2016  
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DOI:10.3969/j.issn.1005-4847.2016.04.003
KeyWord:Schizopygopsis pylzovi;RNA interference;Myostatin;Creatine kinase;Expression
           
AuthorInstitution
孔庆辉 青海大学 农牧学院动物科学系, 西宁
晁燕 青海大学 农牧学院动物科学系, 西宁
夏明哲 青海大学 生态环境工程学院, 西宁
祁得林 青海大学 农牧学院动物科学系, 西宁
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Abstract:
      Objective To investigate the silencing effect of RNA interference on MSTN gene (myostatin, MSTN) expression, and detect the effects on the downstream genes in Schizopygopsis pylzovi.Methods To construct the recombinant adenovirus vector 1P3 (DSP MSTN 273+250+1737) and 1P2 (DSP MSTN 195+1670) for RNA interference of the MSTN gene in Schizopygopsis pylzovi,and to conduct the RNA interference in vivo experiment by injecting the vector into the muscle tissue of Schizopygopsis pylzovi. Real-time PCR and Western blotting were used to evaluate the silencing effects on MSTN gene expression, and to detect the regulatory function of M-CK at gene transcription level after RNA interference of the MSTN gene.Results The result of real-time PCR showed that compared with the HK team (Virus general negative control group) and N team (blank control group), the 1P3 had significant interference effect on the MSTN gene transcription in Schizopygopsis pylzovi (P<0.05), with an inhibition rate of 53.5%, but the 1P2 had no significant interference effect on the MSTN gene transcription. The result of Western blotting was consistent with the results of real-time PCR. At the same time, after the 1P3 interference, the level of MSTN gene transcription was declined, and the level of M-CK gene expression was significantly increased. Conclusions Our results demonstrate that the expression of MSTN gene can be effectively suppressed, and the expression of M-CK gene can be up-regulated through the RNA interference. Therefore, it proves that MSTN gene can inhibit the transcription of M-CK gene in Schizopygopsis pylzovi,and reveals the regulatory role of MSTN gene in the muscle growth and development in the plateau fish Schizopygopsis pylzovi.
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