[Objective]Solute carrier family 11 member 1(SLC11A1)is a major natural resistance candidate gene,which contributes to defense mechanisms of a variety of intracellular bacteria.The SLC11A1 gene promoter sequence of ...[Objective]Solute carrier family 11 member 1(SLC11A1)is a major natural resistance candidate gene,which contributes to defense mechanisms of a variety of intracellular bacteria.The SLC11A1 gene promoter sequence of Xinjiang Brown Cattle,Holstein and Simmental were cloned in the test,and promoter sequence difference was analyzed,in order to provide genetic marker-assisted selection for disease-resistant breeding of dairy cattle.[Method]The Genomic DNA was extracted from whole blood collected from three cattle breeds in Xinjiang,and the 5’ flanking region of SLC11A1 gene was amplified by PCR and sequenced.The sequence was analyzed by bioinformatics software CpGplot,RepeatMasker,TFSEARCH,WWW Signal Scan and dual luciferase assay system.[Result]The SLC11A1 gene promoter sequence of 1 463 bp was confirmed,which had promoter activity.No CpG islands were found on promoter sequence.There were four different sites in SLC11A1 gene promoter sequences between Angus from America and three cattle breeds in Xinjiang.Sequence analysis revealed 12 transcription factor binding sites including Sp1,NF1,RelA-p65,GKLF,and CPBP.In promoter region there was an enhancer region(-734- -740)and two short scattered repetitive elements BOV-tA2,MIR3,as well as repeated DNA element Charlie8.[Conclusion]The SLC11A1 gene promoter sequences of three breeds were obtained,which were different from that of Angus.The paper provided a theoretical basis for further studying the influence of SLC11A1 gene polymorphisms on resistance against intracellular bacteria infection.展开更多
基金Supported by Basic Scientific Research Fund for Public-Interest Scientific Research Institutes in Xinjiang Uygur Autonomous Region(KY2014008)
文摘[Objective]Solute carrier family 11 member 1(SLC11A1)is a major natural resistance candidate gene,which contributes to defense mechanisms of a variety of intracellular bacteria.The SLC11A1 gene promoter sequence of Xinjiang Brown Cattle,Holstein and Simmental were cloned in the test,and promoter sequence difference was analyzed,in order to provide genetic marker-assisted selection for disease-resistant breeding of dairy cattle.[Method]The Genomic DNA was extracted from whole blood collected from three cattle breeds in Xinjiang,and the 5’ flanking region of SLC11A1 gene was amplified by PCR and sequenced.The sequence was analyzed by bioinformatics software CpGplot,RepeatMasker,TFSEARCH,WWW Signal Scan and dual luciferase assay system.[Result]The SLC11A1 gene promoter sequence of 1 463 bp was confirmed,which had promoter activity.No CpG islands were found on promoter sequence.There were four different sites in SLC11A1 gene promoter sequences between Angus from America and three cattle breeds in Xinjiang.Sequence analysis revealed 12 transcription factor binding sites including Sp1,NF1,RelA-p65,GKLF,and CPBP.In promoter region there was an enhancer region(-734- -740)and two short scattered repetitive elements BOV-tA2,MIR3,as well as repeated DNA element Charlie8.[Conclusion]The SLC11A1 gene promoter sequences of three breeds were obtained,which were different from that of Angus.The paper provided a theoretical basis for further studying the influence of SLC11A1 gene polymorphisms on resistance against intracellular bacteria infection.