Cloning and Transcriptional Activity Analysis of Porcine Fatty Acid Binding Protein 4 Promoter

LIU Keke^1,2, ZHONG Jie¹, LI Hongqiang¹, ZHANG Jin², DONGFANG Yang¹*, WU Wenjing²*

( ¹College of Agronomy and Biotechnology, Hebei Normal University of Science and Technology, Qinhuangdao 066000, China; ²College of Biological Chemical Sciences and Engineering, Jiaxing University, Jiaxing 314000, China)

Abstract:

The purpose of this study was to analyze the transcriptional regulation mechanism of porcine FABP4 (fatty acid binding protein 4) gene and obtain porcine adipose tissue specific promoter elements. Using 6-month-old large-white pigs as materials, it was proved that porcine FABP4 was a gene specifically expressed in adipose tissue by fluorescence quantitative PCR and Western blot analysis. The core promoter region of FABP4 was located between –2115 bp—–2066 bp. The truncated fragments 870 bp, 2 914 bp, 5 060 bp upstream of the start codon were cloned, and dual luciferase reporter vectors were constructed naming as pGL3-F-1 Kb/3 Kb/5 Kb. It was found that pGL3-F-3 Kb had the highest activity in adipocytes, but had no activity in non-adipocytes. By co-transfection of transcription factors related to lipid metabolism, it was found that C/EBPα positively regulated FABP4 transcriptional activity, while KLF4 and C/EBPβ negatively regulated the transcriptional activity of FABP4. This study showed that the 3 Kb fragment upstream of the start codon of porcine FABP4 gene had adipose tissue-specific promoter activity, which could be used to construct adipose tissue-specific gene modified pigs.

CSTR: 32200.14.cjcb.2022.02.0007