China Animal Husbandry and Veterinary Medicine ›› 2022, Vol. 49 ›› Issue (7): 2442-2450.doi: 10.16431/j.cnki.1671-7236.2022.07.002

• Biotechnology • Previous Articles     Next Articles

Identification of the Core Promoter Region of MC1R Gene in Silver Fox

LIU Huayun, ZHANG Lei, XU Guili, ZHANG Tianhao, XIE Yuchun, DUAN Lingxin, LIU Zhengzhu, GONG Yuanfang   

  1. Hebei Key Laboratory of Specialty Animal Germplasm Resources Exploration and Innovation, College of Animal Science and Technology, Hebei Normal University of Science & Technology, Qinhuangdao 066004, China
  • Received:2021-10-22 Online:2022-07-05 Published:2022-06-29

Abstract: 【Objective】 This study was aimed to screen the core promoter region of melanocortin 1 receptor (MC1R) gene of silver fox,and provide a theoretical basis for exploring the expression regulatory mechanism of MC1R gene.【Method】 Using genomic DNA of silver fox as template,the sequence of the 5'-UTR region of MC1R gene was obtained by PCR amplification.The active region of the MC1R gene promoter of silver fox was predicted comprehensively using 3 online biology softwares.The deletion fragments of different lengths in 5'-UTR region of MC1R gene were amplified by PCR,which were cloned into pMD19-T vector,and the recombinant plasmid was transfected into melanin B16 cells.The luciferase activities of 10 missing fragments were detected by dual luciferase detection technique.【Result】 The 5'-UTR region sequence of MC1R gene in silver fox was successfully obtained,and ―596/+73 bp might be the active region of the promoter using biology softwares prediction.The detection of dual luciferase activity revealed that the constructed luciferase expression vectors of 10 different deletion fragments all had promoter activity.Among them,pGL3-MC1RP8 (―520/+73 bp) had strong activity,indicating that it was the core promoter region,which was basically consistent with the software prediction results.【Conclusion】 The core promoter region of MC1R gene in silver fox was determined to be ―520/+73 bp,which laid a theoretical foundation for the further study on coat color regulation mechanism of MC1R gene.

Key words: silver fox; MC1R gene; promoter; core region

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