Transcriptional Activity of the FUT1 Gene Promoter Region in Pigs

International Journal of Molecular Sciences

Chen Zi²  Zhengchang Wu²  Jing Wang²  Yongjiu Huo²  Guoqiang Zhu¹  Shenglong Wu² 

[1] College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, Jiangsu, China; E-Mail:;Key Laboratory for Animal Genetics, Breeding, Reproduction and Molecular Design, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, Jiangsu, China; E-Mails:

关键词: pig;    FUT1 gene;    promoter;    transcriptional activity;    luciferase;   

DOI  :  10.3390/ijms141224126

来源: mdpi

【 摘 要 】

This study aims to provide a theoretical basis on the regulatory mechanism of the α-l,2-fucosyltransferase (FUT1) gene in pigs by analyzing the transcriptional activity of its promoter region. On the basis of the previously obtained promoter sequence, primers upstream and downstream of the gene were designed using the restriction endonucleases KpnI and HindIII respectively, and the recombinant plasmids of the pGL3-promoter were constructed by inserting promoter sequences with partially missing regions. The resultant mutants were observed by transient transfection assay into HEK293 cells, and the transcriptional activity of the promoter region was determined by luciferase activity. The 5′-flanking region of the FUT1 gene (−1150 to +50 bp) exhibited promoter activity. The −1150-bp to −849-bp region showed negative regulation of the gene. The recombinant plasmid pGL3-898 showed the strongest luciferase activity, and the activity showed a decreasing trend when the deleted region was increased. Recombinant plasmids were successfully constructed, verified, and the positive and negative regulation areas and core promoter region were detected, providing a deeper insight into the transcriptional regulatory mechanism of the FUT1 gene.

【 授权许可】

CC BY   
© 2013 by the authors; licensee MDPI, Basel, Switzerland