牛ATP5B基因启动子与组织差异性表达研究

doi:10.11843/j.issn.0366-6964.2015.03.005

Abstract

Abstract:

The research was conducted to detect the activities of the bovine ATP5B dual luciferase report gene promoter recombinant plasmid by transfecting into 3T3L-1cells and its relative expression in 16 different tissues.Dual luciferase report gene vectors of bovine ATP5B gene promoter were transfected into 3T3L-1 cells by LPS，then the activities of 7 recombined plasmids were measured，the possible core promoter was further confirmed by bioinformatics software.The RNA of 16 different tissues in Qinchuan cattle was extracted by kit.After reverse transcription，the cDNA was tested，then ATP5B gene real-time quantitative primers and a pair of β-actin internal primers were designed.Finally，the proximal ATP5B gene promoter and the amino acid sequence of different species were analyzed by software.Luciferase activity assay indicated that pATP5B-462 had the highest Luciferase activity compared to pGL3-Basic，sequence analyzing indicated that promoter was in the regions of -547--230 bp，including TATA box，CAAT box，GATA box，v-Myb，deltaE.The analysis of ATP5B gene expression in Qinchuan bull showed that，the expression levels in the hind leg muscle and longissimus muscle were relatively high，in testosterone fat，heart，stomach，rennet，large intestine，small intestine，back fat，kidney and liver were relatively moderate，in reticulum，spleen，lung，caecum and rumen were relatively low.According to the proximal promoter，amino acid sequence alignment and the results of software analysis，ATP5B proximal promoter of bovine，goat,sheep,mice，pigs and human had higher similarity.The similarity of amino acids for ATP5B coding among goat，sheep，mice，human and pig were as high as 99%，98%，96%，96%，91%.This research confirmed the promoter region，got tissues expression profiles of bovine ATP5B gene，the results of software alignment revealed that this is a very conservative gene，this lays a foundation for further research on ATP5B gene transcription regulation and function.

CLC Number:

S823.8

DUAN Mei-yan,ZHAO Zhi-dong，LI An-ning，LI Shi-jun,WANG Ming-ming,ZAN Lin-sen. Analysis of Bovine ATP5B Gene Promoter and Tissue Expression[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(3): 382-387.

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Analysis of Bovine ATP5B Gene Promoter and Tissue Expression

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