昆明种小鼠线粒体转录终止因子1基因的组织表达谱分析
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摘要
目的:探索Mterf1基因mRNA和蛋白质在昆明种小鼠大脑、心脏、肝脏、脾脏、肺脏、肾脏、胰腺、骨骼肌、膀胱和卵巢共10种组织的表达情况。方法:采用Real-time RT-PCR检测Mterf1 mRNA在小鼠多个组织中的表达情况;采用Western Blot法检测Mterf1蛋白在小鼠多种组织中的表达情况。结果:Mterf1基因在昆明种小鼠大脑、心脏、骨骼肌和膀胱组织中表达量较高,其次是在肝脏、肾脏和脾脏组织,而在肺脏、胰腺和卵巢组织中表达量相对较低。结论:Mterf1基因在昆明种小鼠中的表达具有组织特异性,为后续研究Mterf1基因在小鼠体内的功能奠定基础。
Objective: To analyze the tissue expression profile of mouse mitochondrial transcription termination factor 1(Mterf1 gene in 10 different tissues of Kunming mice. Methods: Real-time RT-PCR was performed to study the mRNA expression level of Mterf1 gene in different tissues of mice, and Western Blot was preformed to study the protein expression level of Mterf1 protein in different tissues of mice. Results: Compared with the rest of the tissues, the expression level of Mterf1 gene was highest in the brain,heart, skeletal muscle and bladder, secondly in the liver, kidney and spleen, but relatively lower in lung, pancreas and ovary.Conclusion: The expression of Mterf1 gene is tissue-specific in Kunming mice, which will provide a basis for the study of Mterf1 gene function in vivo.
Objective: To analyze the tissue expression profile of mouse mitochondrial transcription termination factor 1(Mterf1 gene in 10 different tissues of Kunming mice. Methods: Real-time RT-PCR was performed to study the mRNA expression level of Mterf1 gene in different tissues of mice, and Western Blot was preformed to study the protein expression level of Mterf1 protein in different tissues of mice. Results: Compared with the rest of the tissues, the expression level of Mterf1 gene was highest in the brain,heart, skeletal muscle and bladder, secondly in the liver, kidney and spleen, but relatively lower in lung, pancreas and ovary.Conclusion: The expression of Mterf1 gene is tissue-specific in Kunming mice, which will provide a basis for the study of Mterf1 gene function in vivo.
引文
[1]黄淑颜,丘式浚,陈家逸,等.线粒体转录终止因子(mTERF)蛋白家族的研究进展[J].生命科学研究,2016,20(5):455-459.
[2]LINDER T,PARK C B,ASIN-CAYUELA J,et al.A family of putative transcription termination factors shared amongst metazoans and plants[J].Current Genetics,2005,48(4):265-269.
[3]KRUSE B,NARASIMHAN N,ATTARDI G.Termination of transcription in human mitochondria:identification and purification of a DNA binding protein factor that promotes termination[J].Cell,1989,58(2):391-397.
[4]CHEN G,DAI J,TAN S,et al.MTERF1 regulates the oxidative phosphorylation activity and cell proliferation in HeLa cells[J].Acta Biochim Biophys Sin(Shanghai),2014,46(6):512-521.
[5]FERNANDEZ-SILVA P,MARTINEZ-AZORIN F,MICOLV,et al.The human mitochondrial transcription termination factor(mTERF)is a multizipper protein but binds to DNA as a monomer,with evidence pointing to intramolecular leucine zipper interactions[J].EMBO J,1997,16(5):1066-1079.
[6]DAGA A,MICOL V,HESS D,et al.Molecular characterizaiton of the transcirption termination factor from human mitochondira[J].J Biol Chem,1993,268(11):8123-8130.
[7]HYV?RINEN A K,KUMANTO M K,MARJAVAARA SK,et al.Effects on mitochondrial transcription of manipulating mTERF protein levels in cultured human HEK293cells[J].BMC Mol Biol,2010,11(9):72-78.
[8]熊伟,张海洋,杨红娟,等.哺乳动物细胞线粒体基因组的转录及调控机制研究进展[J].大理学院学报,2014,13(8):26-30.
[9]郭俊良,胡靖扬,高顺玉.昆明小鼠不同组织RNA提取方法的比较[J].楚雄师范学院学报,2011,26(3):84-88.
[10]伍立夫,林春龙.线粒体转录终止因子家族的研究现状及进展[J].医学综述,2015,21(21):3870-3873.
[11]ROBERTI M,POLOSA P L,BRUNI F,et al.MTERF factors:a multifunction protein family[J].Biomol Concepts,2010,1(2):215-224.
[12]MARTIN M,CHO J,CESARE A J,et al.Termination factor-mediated DNA loop between termination and initiation sites drives mitochondrial rRNA synthesis[J].Cell,2005,123(7):1227-1240.
[13]SHI Y,POSSE V,ZHU X,et al.Mitochondrial transcription termination factor 1 directs polar replication fork pausing[J].Nucleic Acids Res,2016,44(12):5732-5742.
[14]VAN DEN OUWELAND J M,LEMKES H H,RUITEN-BEEK W,et al.Mutation in mitochondrial t RNA(Leu)(UUR)gene in a large pedigree with maternally transmitted type II diabetes mellitus and deafness[J].Nat Genet,1992,1(5):368-371.
[15]HESS J F,PARISI M A,BENNETT J L,et al.Impairment of mitochondrial transcription termination by a point mutation associated with the MELAS subgroup of mitochondrial encephalomyopathies[J].Nature,1991,351(6323):236-239.
[16]LI X,ZHANG L S,GUAN M X.Cloning and characterization of mouse mTERF encoding a mitochondrial transcriptional termination factor[J].Biochem Biophys Res Commun,2005,326(2):505-510.
[17]熊伟,余敏,左绍远.线粒体转录终止因子蛋白家族在线粒体基因表达中的调节作用[J].中国生物化学与分子生物学报,2015,31(3):223-231.
[18]TERZIOGLU M,RUZZENENTE B,HARMEL J,et al.MTERF1 binds mtDNA to prevent transcriptional interference at the light-strand promoter but is dispensable for rRNA gene transcription regulation[J].Cell Metab,2013,17(4):618-626.
[19]左谦益,宁磊.昆明小鼠生长发育指标及繁殖性能测定[J].中国比较医学杂志,2001,11(4):199-202.
[20]朱大诚,罗小泉,王艳辉.40日龄昆明小鼠主要生物学特性标准化研究[J].江西中医学院学报,2008,20(2):63-65.
[21]赖青鸟,畅荣妮,袁广之,等.小鼠载脂蛋白E基因的组织表达谱分析[J].广西医科大学学报,2013,30(6):847-849.
[2]LINDER T,PARK C B,ASIN-CAYUELA J,et al.A family of putative transcription termination factors shared amongst metazoans and plants[J].Current Genetics,2005,48(4):265-269.
[3]KRUSE B,NARASIMHAN N,ATTARDI G.Termination of transcription in human mitochondria:identification and purification of a DNA binding protein factor that promotes termination[J].Cell,1989,58(2):391-397.
[4]CHEN G,DAI J,TAN S,et al.MTERF1 regulates the oxidative phosphorylation activity and cell proliferation in HeLa cells[J].Acta Biochim Biophys Sin(Shanghai),2014,46(6):512-521.
[5]FERNANDEZ-SILVA P,MARTINEZ-AZORIN F,MICOLV,et al.The human mitochondrial transcription termination factor(mTERF)is a multizipper protein but binds to DNA as a monomer,with evidence pointing to intramolecular leucine zipper interactions[J].EMBO J,1997,16(5):1066-1079.
[6]DAGA A,MICOL V,HESS D,et al.Molecular characterizaiton of the transcirption termination factor from human mitochondira[J].J Biol Chem,1993,268(11):8123-8130.
[7]HYV?RINEN A K,KUMANTO M K,MARJAVAARA SK,et al.Effects on mitochondrial transcription of manipulating mTERF protein levels in cultured human HEK293cells[J].BMC Mol Biol,2010,11(9):72-78.
[8]熊伟,张海洋,杨红娟,等.哺乳动物细胞线粒体基因组的转录及调控机制研究进展[J].大理学院学报,2014,13(8):26-30.
[9]郭俊良,胡靖扬,高顺玉.昆明小鼠不同组织RNA提取方法的比较[J].楚雄师范学院学报,2011,26(3):84-88.
[10]伍立夫,林春龙.线粒体转录终止因子家族的研究现状及进展[J].医学综述,2015,21(21):3870-3873.
[11]ROBERTI M,POLOSA P L,BRUNI F,et al.MTERF factors:a multifunction protein family[J].Biomol Concepts,2010,1(2):215-224.
[12]MARTIN M,CHO J,CESARE A J,et al.Termination factor-mediated DNA loop between termination and initiation sites drives mitochondrial rRNA synthesis[J].Cell,2005,123(7):1227-1240.
[13]SHI Y,POSSE V,ZHU X,et al.Mitochondrial transcription termination factor 1 directs polar replication fork pausing[J].Nucleic Acids Res,2016,44(12):5732-5742.
[14]VAN DEN OUWELAND J M,LEMKES H H,RUITEN-BEEK W,et al.Mutation in mitochondrial t RNA(Leu)(UUR)gene in a large pedigree with maternally transmitted type II diabetes mellitus and deafness[J].Nat Genet,1992,1(5):368-371.
[15]HESS J F,PARISI M A,BENNETT J L,et al.Impairment of mitochondrial transcription termination by a point mutation associated with the MELAS subgroup of mitochondrial encephalomyopathies[J].Nature,1991,351(6323):236-239.
[16]LI X,ZHANG L S,GUAN M X.Cloning and characterization of mouse mTERF encoding a mitochondrial transcriptional termination factor[J].Biochem Biophys Res Commun,2005,326(2):505-510.
[17]熊伟,余敏,左绍远.线粒体转录终止因子蛋白家族在线粒体基因表达中的调节作用[J].中国生物化学与分子生物学报,2015,31(3):223-231.
[18]TERZIOGLU M,RUZZENENTE B,HARMEL J,et al.MTERF1 binds mtDNA to prevent transcriptional interference at the light-strand promoter but is dispensable for rRNA gene transcription regulation[J].Cell Metab,2013,17(4):618-626.
[19]左谦益,宁磊.昆明小鼠生长发育指标及繁殖性能测定[J].中国比较医学杂志,2001,11(4):199-202.
[20]朱大诚,罗小泉,王艳辉.40日龄昆明小鼠主要生物学特性标准化研究[J].江西中医学院学报,2008,20(2):63-65.
[21]赖青鸟,畅荣妮,袁广之,等.小鼠载脂蛋白E基因的组织表达谱分析[J].广西医科大学学报,2013,30(6):847-849.
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