The Effects Of TSMR1 On The Heart Development Candidate Gene Promoter Activity In Drosophila

TSMR1 is a candidate gene for the development of human heart in our laboratory,and their function has not been reported yet.In order to study the role of the human gene TSMR1 in the development of heart,we have shown that TSMR1(human homologous TSMR1)is a candidate gene for heart development.The embryonic development of Drosophila melanogaster affects the arrangement of the myocardial cells and may also affect the function of the adult Drosophila heart.In order to study the role of TSMR1 in the development of heart,and to study the effect of it function on the activity of cardiac gene promoter.I extracted the adult wild type Drosophila DNA,primers were designed to clone the Drosophila heart development related genes(Eve,Zfh-1,Odd,Prc,Svp,Smox),and cloned into p GL3-basic luciferase reporter vector to construct the expression plasmid p GL3-Eve,p GL3-Zfh1,p GL3-Odd,p GL3-Prc,p GL3-Svp,p GL3-Smox.Plasmids were co-transfected with PCMV-myc-TSMR1 into HEK293 cells,meanwhile p GL3-basic,PCMV-myc and those promoter reporter plasmid were transfected into HEK293 cells individually.p RL-TK as an internal reference,luciferase reporter gene detection system was established.The promoter activity of cardiac development related genes was identified by luciferase reporter assay system,and the data were analyzed by independent sample t test.The results showed that the TSMR1 gene could enhance the transcriptional activity of the promoter.In order to further study the specific site of the promoter region,the fragment of the cardiac gene promoter region was analyzed and divided into 5-6 segment fragment with a length of about 500 bp.Luciferase reporter assay was used to detect promoter activity as same.The results show that the core promoter region of Svp is-465 — +40bp;the core promoter region of Prc is-1426 —-900bp;the core promoter region of Smox is-1205 —-646bp;the core promoter region of Zfh1 is-2646 —-2127 bp.In summary,this study identified the regulatory role of Drosophila TSMR1 on cardiac gene promoter activity,which laid the foundation for further study of the regulatory mechanism of TSMR1 in Drosophila heart development.