徐淮山羊c-Myc基因启动子的克隆及其功能的初步分析

doi:10.11843/j.issn.0366-6964.2014.04.004

Abstract

Abstract:

The aim of this study was to determine and find the promoter core regulatory regions of Xuhuai goat’s c-Myc gene，and to investigate c-Myc gene regulation mechanism.According to the sequence of ovis aries c-Myc gene published in GenBank，the primers were designed and the different deletion fragments of 5′ flanking region of c-Myc gene promoter were amplified，and then were cloned into pEGFP-N1 and PGL3-Basic，then were transfected into gFF，COS7 and P19 cells and treated with TSA and NFAT1.SP1 transcription factor binding sites in the region of -402--249 bp was mutated，and the activity of dual-luciferase reporter gene was detected.The study showed that the region containing -1 334-+1 bp of c-Myc promoter had the maximal promoter activities，and sequence of -402-+1 bp had the basal promoter activities，further study showed that there were positive (the region of -1 334--971 bp，-587--147 bp) and negative (the region of -1 976--1 334 bp，-971--587 bp) regulatory domains，respectively.TSA and NFAT1 significantly enhanced the activity of c-Myc promoter.The activity was declined by the point mutation of SP1 binding site.By constructing the recombinant expression vectors containing different fragments of c-Myc gene promoter and comparing their activity，the core regulatory region of c-Myc gene promoter was identified.SP1 is an important regulatory element of c-Myc promoter core region，which lay a foundation for the further research on the mechanism of regulation and expression of c-Myc gene.

CLC Number:

S827
S813.3

WEI Guang-hui，ZUO Qi-sheng，LI Dong，ZHANG Ya-ni，LIU Zhi-yong，ZHU Rui，QIU Feng-long，LI Bi-chun. Cloning and Preliminary Function Analysis of Xuhuai Goat c-Myc Promoter[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(4): 533-540.

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Cloning and Preliminary Function Analysis of Xuhuai Goat c-Myc Promoter

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