Functional analysis of the promoter of the dmrt1 gene in Chinese tongue sole,Cynoglossus semilaevis
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摘要
The dmrtl(doublesex and mab-3 related transcription factor 1)gene is considered to play an essential role in testis differentiation and development among metazoan species.As an economically important marine fish in China,Chinese tongue sole(Cynoglossus semilaevis)possess a ZW/ZZ sexdetermining sy stem,and the females usually grow two to four times bigger than males.Previous studies have confirmed that Z-linked dmrtl gene is the male determining gene in C.semilaevis and TALENs-mediated knocking-out individuals showed increased growth phenotype.In order to investigate the function of dmrtl regulatory regions and its potential application,we have cloned the promoter of C.semilaevis dmrtl.The acquired sequence consisted of a 5'-upstream sequence and a partial exonl;functional analysis showed that there were two positive regulatory regions in the promoter,as well as one negative region and one neutral region.These positive regions were then combined to obtain several newly constructed promoters.Both in vitro and in vivo analysis confirmed one of them(dmrtl-Δ3+Δ4)showed elevated promoting activity compared to the full-length promoter and could drive exogenous gene expression in tongue sole gonads.Our results will provide useful information for understanding the regulatory mechanism of dmrtl gene during C.semilaevis sex determination,and the endogenous promoter will facilitate the transgenic research for analyzing sex-and growth-related genes.
The dmrtl(doublesex and mab-3 related transcription factor 1)gene is considered to play an essential role in testis differentiation and development among metazoan species.As an economically important marine fish in China,Chinese tongue sole(Cynoglossus semilaevis)possess a ZW/ZZ sexdetermining sy stem,and the females usually grow two to four times bigger than males.Previous studies have confirmed that Z-linked dmrtl gene is the male determining gene in C.semilaevis and TALENs-mediated knocking-out individuals showed increased growth phenotype.In order to investigate the function of dmrtl regulatory regions and its potential application,we have cloned the promoter of C.semilaevis dmrtl.The acquired sequence consisted of a 5'-upstream sequence and a partial exonl;functional analysis showed that there were two positive regulatory regions in the promoter,as well as one negative region and one neutral region.These positive regions were then combined to obtain several newly constructed promoters.Both in vitro and in vivo analysis confirmed one of them(dmrtl-Δ3+Δ4)showed elevated promoting activity compared to the full-length promoter and could drive exogenous gene expression in tongue sole gonads.Our results will provide useful information for understanding the regulatory mechanism of dmrtl gene during C.semilaevis sex determination,and the endogenous promoter will facilitate the transgenic research for analyzing sex-and growth-related genes.
The dmrtl(doublesex and mab-3 related transcription factor 1)gene is considered to play an essential role in testis differentiation and development among metazoan species.As an economically important marine fish in China,Chinese tongue sole(Cynoglossus semilaevis)possess a ZW/ZZ sexdetermining sy stem,and the females usually grow two to four times bigger than males.Previous studies have confirmed that Z-linked dmrtl gene is the male determining gene in C.semilaevis and TALENs-mediated knocking-out individuals showed increased growth phenotype.In order to investigate the function of dmrtl regulatory regions and its potential application,we have cloned the promoter of C.semilaevis dmrtl.The acquired sequence consisted of a 5'-upstream sequence and a partial exonl;functional analysis showed that there were two positive regulatory regions in the promoter,as well as one negative region and one neutral region.These positive regions were then combined to obtain several newly constructed promoters.Both in vitro and in vivo analysis confirmed one of them(dmrtl-Δ3+Δ4)showed elevated promoting activity compared to the full-length promoter and could drive exogenous gene expression in tongue sole gonads.Our results will provide useful information for understanding the regulatory mechanism of dmrtl gene during C.semilaevis sex determination,and the endogenous promoter will facilitate the transgenic research for analyzing sex-and growth-related genes.
引文
Akerfelt M,Morimoto R I,Sistonen L.2010.Heat shock factors:integrators of cell stress,development and lifespan.Nat Rev.Mol.Cell.Biol,11(8):545-555.
Chakraborty T,Zhou L Y,Chaudhari A,Iguchi T,Nagahama Y.2016.Dmy initiates masculinity by altering Gsdfl Sox9a21Rspo 1 expression in medaka(Oryzias latipes).Sci.Rep.,6:19 480.
Chen S L,Tian Y S,Yang J F,Shao C W,Ji X S,Zhai J M,Liao X L,Zhuang Z M,Su P Z,Xu J Y,Sha Z X,Wu P F,Wang N.2009.Artificial gynogenesis and sex determination in half-smooth tongue sole(Cynoglossus semilaevis).Mar.Biotechnol, 11(2):243-25 1.
Chen S L,Zhang G J,Shao C W,Huang Q F,Liu G,Zhang P,Song W T,An N,Chalopin D,Volff J N,Hong Y H,Li Q Y,Sha Z X,Zhou H L,Xie M S,Yu Q L,Liu Y,Xiang H,Wang N,Wu K,Yang C G,Zhou Q,Liao X L,Yang L F,Hu Q M,Zhang J L,Meng L,Jin L J,Tian Y S,Lian J M,Yang J F,Miao G D,Liu S S,Liang Z,Yan F,Li Y Z,Sun B,Zhang H,Zhang J,Zhu Y,Du M,Zhao Y W,Schartl M,Tang Q S,Wang J.2014.Whole-genome sequence of a flatfish provides insights into ZW sex chromosome evolution and adaptation to a benthic lifestyle.Nat.Genet.,46(3):253-260.
Cook T,Gebelein B,Urrutia R.1999.Spl and its likes:biochemical and functional predictions for a growing family of zinc finger transcription factors.Ann.N.Y.Acad.Sci.,880(1):94-102.
Cui Z K,Liu Y,Wang W W,Wang Q,Zhang N,Lin F,Wang N,Shao C W,Dong Z D,Li Y Z,Yang Y M,Hu M Z,Li H L,Gao F T,Wei Z F,Meng L,Liu Y,Wei M,Zhu Y,Guo H,Cheng C H K,Schartl M,Chen S L.2017.Genome editing reveals dmrt1 as an essential male sex-determining gene in Chinese tongue sole(Cynoglossus semilaevis).Sci.Rep.,7:42 213.
Friedenreich H,Schartl M.1990.Transient expression directed by homologus and heterologous prometer and enhancer sequences in fish cells.Nucleic Acids Res.,18(11):3 299-3 305.
Gao S,Zhang T,Zhou X,Zhao Y,Li Q,Guo Y Q,Cheng H H.2005.Molecular cloning,expression of Sox5 and its down-regulation of Dmrtl transcription in zebrafish.J.Exp.Zool. B Mol Dev.Evol.,304B(5):476-483.
Guan G J,Kobayashi T,Nagahama Y.2000.Sexually dimorphic expression of twotpes of DM.(Doublesexl Ma b-3)-domain genes in a teleo st fish,the Tilapia(Oreochromis niloticus).Biochem.Biophys.Res.Commun.,272(3):662-666.
Gubbay J,Collignon J,Koopman P,Capel B,Economou A,Münsterberg A,Vivian N,Goodfellow P,Lovell-Badge R.1990.A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes.Nature,346(6281):245-250.
Ji X S,Liu H W,Chen S L,Jiang Y L,Tian Y S.2011.Growth differences and dimorphic expression of growth hormone(GH)in female and male Cynoglossus semilaevis after male sexual maturation.Mar.Genomics,4(1):9-16.
Jorgensen A,Morthorst J E,Andersen O,Rasmussen L J,Bjerregaard P.2008.Expression profiles for six zebrafish genes during gonadal sex differentiation.Reprod.Biol.Endocrinol.,6(1):25.
Kfoury N,Kapatos G.2009.Identification of neuronal target genes for CCAAT/enhancer binding proteins.Mol.Cell.Neurosci.,40(3):3 1 3-327.
Kobayashi T,Matsuda M,Kajiura-Kobayashi H,Suzuki A,Saito N,Nakamoto M,Shibata N,Nagahama Y.2004.Two DM domain genes,DMY and DMRT1.involved in testicular differentiation and development in the medaka,Oryzias latipes.Dev.Dyn.,231(3):518-526.
Koopman P.2009.Sex determination:the power of DMRTl.Trends Genet.,25(11):479-48 1.
Lambeth L S,Raymond C S,Roeszler K N,Kuroiwa A,Nakata T,Zarkower D,Smith C A.2014.Over-expression of DMRT1 induces the male pathway in embryonic chicken gonads.Dev.Biol.,389(2):160-172.
Lei N,Heckert L L.2002.Spl and Egrl regulate transcription of the Dmrtl gene in Sertoli cells.Biol.Reprod.,66(3):675-684.
Lei N,Karpova T,Hornbaker K I,Rice D A,Heckert L L.2009.Distinct transcriptional mechanisms direct expression of the rat Dmrtl promoter in sertoli cells and germ cells of transgenic mice.Biol.Reprod.,81(1):118-125.
Luo D J,Liu Y,Chen J,Xia X Q,Cao M X,Cheng B,Wang X J,Gong W M,Qiu C,Zhang Y S,Ki Cheng,C H,Zhu Z Y,Hu W.2015.Direct production of XY~(DMY-)sex reversal female medaka(Oryzias latipes)by embryo micro inj ection of TALENs.Sci.Rep.,5:14 057.
Marchand O,Govoroun M,D'Cotta H,McMeel O,Lareyre J J,Bernot A,Laudet V,Guiguen Y.2000.DMRT1expression during gonadal differentiation andspermatogenesis in the rainbow trout,Oncorhynchus mykiss.Biochim.Biophys.Acta,1493(1-2):1 80-1 87.
Marin M,Karis A,Visser P,Grosveld F,Philipsen S.1997.Transcription factor Spl is essential for early embryonic development but dispensable for cell growth and differentiation.Cell,89(4):619-628.
Matsuda M,Nagahama Y,Shinomiya A,Sato T,Matsuda C,Kobayashi T,Morrey C E,Shibata N,Asakawa S,Shimizu N,Hori H,Hamaguchi S,Sakaizumi M.2002.DMYis,a Y-specific DM-domain gene required for male development in the medaka fish.Nature,417(6888):5 5 9-563.
Matsuda M,Shinomiya A,Kinoshita M,Suzuki A,Kobayashi T,Paul-Prasanth B,Lau E L,Hamaguchi S,Sakaizumi M,Nagahama Y.2007.DMYgene induces male development in genetically female(XX)medaka fish.Proc.Natl.Acad.Sci.USA,104(10):3 865-3 870.
Mawaribuchi S,Musashijima M,Wada M,Izutsu Y,Kurakata E,Park M K,Takamatsu N,Ito M.2017.Molecular evolution of two distinct dmrtl promoters for germ and somatic cells in vertebrate gonads.Mol.Biol.Evol.,34(3):724-733.
Nanda I,Kondo M,Hornung U,Asakawa S,Winkler C,Shimizu A,Shan Z H,Haaf T,Shimizu N,Shima A,Schmid M,Schartl M.2002.A duplicated copy of DMRT1in the sex-determining region of the Y chromosome of the medaka,Oryzias latipes.Proc.Natl.Acad.Sci.USA,99(18):11 778-11 783.
Paul-Prasanth B,Matsuda M,Lau E L,Suzuki A,Sakai F,Kobayashi T,Nagahama Y.2006.Knock-down of DMY initiates female pathway in the genetic male medaka,Oryzias latipes.Biochem.Biophys.Res.Commun.,351(4):815-819.
Quitschke W W,Lin Z Y,DePonti-Zilli L,Paterson B M.1989.The beta actin promoter.High levels of transcription depend upon a CCAAT binding factor.J.Biol.Chem.,264(16):9 539-9 546.
Raymond C S,Murphy M W,O'Sullivan M G,Bardwell V J,Zarkower D.2000.Dmrtl,a gene related to worm and fly sexual regulators,is required for mammalian testis differentiation.Genes Dev.,14(20):2 587-2 595.
Raymond C S,Parker E D,Kettlewell J R,Brown L G,Page D C,Kusz K,Jaruzelska J,Reinberg Y,Flej ter W L,Bardwell V J,Hirsch B,Zarkower D.1999.A region of human chromosome 9p required for testis development contains two genes related to known sexual regulators.Hum.Mol.Genet.,8(6):989-996.
Raymond C S,Shamu C E,Shen M M,Seifert K J,Hirsch B,Hodgkin J,Zarkower D.1998.Evidence for evolutionary conservation of sex-determining genes.Nature,391(6668):691-695.
Shao C W,Li Q Y,Chen S L,Zhang P,Lian J M,Hu Q M,Sun B,Jin L J,Liu S S,Wang Z J,Zhao H M,Jin Z H,LiangZ,Li YZ,Zheng Q M,Zhang Y,Wang J,Zhang G J.2014.Epigenetic modification and inheritance in sexual reversal of fish.Genome Res.,24(4):604-615.
Shetty S,Kirby P,Zarkower D,Graves JA M.2002.DMRT1in a ratite bird:evidence for a role in sex determination and discovery of a putative regulatory element.Cytogenet.Genome Res.,99(1-4):245-25 1.
Sinclair A H,Berta P,Palmer M S,Hawkins J R,Griffiths B L,Smith M J,Foster J W,Frischauf A M,Lovell-Badge R,Goodfellow P N.1990.A gene from the human sexdetermining region encodes a protein with homology to a conserved DNA-binding motif.Nature,346(6281):240-244.
Slomiany B A,D'Arigo K L,Kelly M M,Kurtz D T.2000.C/EBPa inhibits cell growth via direct repression of E2FDP-mediated transcription.Mol.Cell.Biol.,20(16):5 986-5 997.
Smith C A,McClive P J,Western P S,Reed K J,Sinclair A H.1999.Conservation of a sex-determining gene.Nature,402(6762):601-602.
Smith C A,Roeszler K N,Ohnesorg T,Cummins D M,Farlie P G,Doran T J,Sinclair A H.2009.The avian Z-linked gene DMRT1 is required for male sex determination in the chicken.Nature,461(7261):267-271.
Sunilkumar G,Connell J P,Smith C W,Reddy A S,Rathore K S.2002.Cotton a-globulin promoter:isolation and functional characterization in transgenic cotton,Arabidopsis,and tobacco.Transgenic Res.,11(4):347-359.
Topilko P,Schneider-Maunoury S,Levi G,Trembleau A,Gourdji D,Driancourt M A,Rao C V,Chamay P.1998.Multiple pituitary and ovarian defects in Krox-24(NGFIA,Egr-1)-targeted mice.Mol Endocrinol.,12(1):107-122.
Tourtellotte W G,Nagarajan R,Auyeung A,Mueller C,Milbrandt J. 1999.Infertility associated with incomplete spermatogenic arrest and oligozoospermia in Egr4-deficient mice.Development,126(22):5 061-5 071.
Tsunoda T,Takagi T.1999.Estimating transcription factor bindability on DNA.Bioinformatics,15(7-8):622-630.
Xia W,Zhou L,Yao B,Li C J,Gui J F.2007.Differential and spermatogenic cell-specific expression of DMRT1 during sex reversal in protogynous hermaphroditic groupers.Mol.Cell.Endocrinol.,263(1-2):1 56-1 72.
Yo shimoto S,Ito M.2011.A ZZ/ZW-type sex determination in Xenopus laevis.FEBS J.,278(7):1 020-1 026.
Zhao L,Svingen T,Ng E T,Koopman P.2015.Female-to-male sex reversal in mice caused by transgenic overexpression of Dmrtl.Development,142(6):1 083-1 088.
Zhou L Q,Yang A G,Liu X Z,Du W,Zhuang Z M.2005.The karyotype of the tonguefish Cynoglossus semilaevis.J.Fish.Sci.China,29(3):417-41 9.(in Chinese with English abstract)
Chakraborty T,Zhou L Y,Chaudhari A,Iguchi T,Nagahama Y.2016.Dmy initiates masculinity by altering Gsdfl Sox9a21Rspo 1 expression in medaka(Oryzias latipes).Sci.Rep.,6:19 480.
Chen S L,Tian Y S,Yang J F,Shao C W,Ji X S,Zhai J M,Liao X L,Zhuang Z M,Su P Z,Xu J Y,Sha Z X,Wu P F,Wang N.2009.Artificial gynogenesis and sex determination in half-smooth tongue sole(Cynoglossus semilaevis).Mar.Biotechnol, 11(2):243-25 1.
Chen S L,Zhang G J,Shao C W,Huang Q F,Liu G,Zhang P,Song W T,An N,Chalopin D,Volff J N,Hong Y H,Li Q Y,Sha Z X,Zhou H L,Xie M S,Yu Q L,Liu Y,Xiang H,Wang N,Wu K,Yang C G,Zhou Q,Liao X L,Yang L F,Hu Q M,Zhang J L,Meng L,Jin L J,Tian Y S,Lian J M,Yang J F,Miao G D,Liu S S,Liang Z,Yan F,Li Y Z,Sun B,Zhang H,Zhang J,Zhu Y,Du M,Zhao Y W,Schartl M,Tang Q S,Wang J.2014.Whole-genome sequence of a flatfish provides insights into ZW sex chromosome evolution and adaptation to a benthic lifestyle.Nat.Genet.,46(3):253-260.
Cook T,Gebelein B,Urrutia R.1999.Spl and its likes:biochemical and functional predictions for a growing family of zinc finger transcription factors.Ann.N.Y.Acad.Sci.,880(1):94-102.
Cui Z K,Liu Y,Wang W W,Wang Q,Zhang N,Lin F,Wang N,Shao C W,Dong Z D,Li Y Z,Yang Y M,Hu M Z,Li H L,Gao F T,Wei Z F,Meng L,Liu Y,Wei M,Zhu Y,Guo H,Cheng C H K,Schartl M,Chen S L.2017.Genome editing reveals dmrt1 as an essential male sex-determining gene in Chinese tongue sole(Cynoglossus semilaevis).Sci.Rep.,7:42 213.
Friedenreich H,Schartl M.1990.Transient expression directed by homologus and heterologous prometer and enhancer sequences in fish cells.Nucleic Acids Res.,18(11):3 299-3 305.
Gao S,Zhang T,Zhou X,Zhao Y,Li Q,Guo Y Q,Cheng H H.2005.Molecular cloning,expression of Sox5 and its down-regulation of Dmrtl transcription in zebrafish.J.Exp.Zool. B Mol Dev.Evol.,304B(5):476-483.
Guan G J,Kobayashi T,Nagahama Y.2000.Sexually dimorphic expression of twotpes of DM.(Doublesexl Ma b-3)-domain genes in a teleo st fish,the Tilapia(Oreochromis niloticus).Biochem.Biophys.Res.Commun.,272(3):662-666.
Gubbay J,Collignon J,Koopman P,Capel B,Economou A,Münsterberg A,Vivian N,Goodfellow P,Lovell-Badge R.1990.A gene mapping to the sex-determining region of the mouse Y chromosome is a member of a novel family of embryonically expressed genes.Nature,346(6281):245-250.
Ji X S,Liu H W,Chen S L,Jiang Y L,Tian Y S.2011.Growth differences and dimorphic expression of growth hormone(GH)in female and male Cynoglossus semilaevis after male sexual maturation.Mar.Genomics,4(1):9-16.
Jorgensen A,Morthorst J E,Andersen O,Rasmussen L J,Bjerregaard P.2008.Expression profiles for six zebrafish genes during gonadal sex differentiation.Reprod.Biol.Endocrinol.,6(1):25.
Kfoury N,Kapatos G.2009.Identification of neuronal target genes for CCAAT/enhancer binding proteins.Mol.Cell.Neurosci.,40(3):3 1 3-327.
Kobayashi T,Matsuda M,Kajiura-Kobayashi H,Suzuki A,Saito N,Nakamoto M,Shibata N,Nagahama Y.2004.Two DM domain genes,DMY and DMRT1.involved in testicular differentiation and development in the medaka,Oryzias latipes.Dev.Dyn.,231(3):518-526.
Koopman P.2009.Sex determination:the power of DMRTl.Trends Genet.,25(11):479-48 1.
Lambeth L S,Raymond C S,Roeszler K N,Kuroiwa A,Nakata T,Zarkower D,Smith C A.2014.Over-expression of DMRT1 induces the male pathway in embryonic chicken gonads.Dev.Biol.,389(2):160-172.
Lei N,Heckert L L.2002.Spl and Egrl regulate transcription of the Dmrtl gene in Sertoli cells.Biol.Reprod.,66(3):675-684.
Lei N,Karpova T,Hornbaker K I,Rice D A,Heckert L L.2009.Distinct transcriptional mechanisms direct expression of the rat Dmrtl promoter in sertoli cells and germ cells of transgenic mice.Biol.Reprod.,81(1):118-125.
Luo D J,Liu Y,Chen J,Xia X Q,Cao M X,Cheng B,Wang X J,Gong W M,Qiu C,Zhang Y S,Ki Cheng,C H,Zhu Z Y,Hu W.2015.Direct production of XY~(DMY-)sex reversal female medaka(Oryzias latipes)by embryo micro inj ection of TALENs.Sci.Rep.,5:14 057.
Marchand O,Govoroun M,D'Cotta H,McMeel O,Lareyre J J,Bernot A,Laudet V,Guiguen Y.2000.DMRT1expression during gonadal differentiation andspermatogenesis in the rainbow trout,Oncorhynchus mykiss.Biochim.Biophys.Acta,1493(1-2):1 80-1 87.
Marin M,Karis A,Visser P,Grosveld F,Philipsen S.1997.Transcription factor Spl is essential for early embryonic development but dispensable for cell growth and differentiation.Cell,89(4):619-628.
Matsuda M,Nagahama Y,Shinomiya A,Sato T,Matsuda C,Kobayashi T,Morrey C E,Shibata N,Asakawa S,Shimizu N,Hori H,Hamaguchi S,Sakaizumi M.2002.DMYis,a Y-specific DM-domain gene required for male development in the medaka fish.Nature,417(6888):5 5 9-563.
Matsuda M,Shinomiya A,Kinoshita M,Suzuki A,Kobayashi T,Paul-Prasanth B,Lau E L,Hamaguchi S,Sakaizumi M,Nagahama Y.2007.DMYgene induces male development in genetically female(XX)medaka fish.Proc.Natl.Acad.Sci.USA,104(10):3 865-3 870.
Mawaribuchi S,Musashijima M,Wada M,Izutsu Y,Kurakata E,Park M K,Takamatsu N,Ito M.2017.Molecular evolution of two distinct dmrtl promoters for germ and somatic cells in vertebrate gonads.Mol.Biol.Evol.,34(3):724-733.
Nanda I,Kondo M,Hornung U,Asakawa S,Winkler C,Shimizu A,Shan Z H,Haaf T,Shimizu N,Shima A,Schmid M,Schartl M.2002.A duplicated copy of DMRT1in the sex-determining region of the Y chromosome of the medaka,Oryzias latipes.Proc.Natl.Acad.Sci.USA,99(18):11 778-11 783.
Paul-Prasanth B,Matsuda M,Lau E L,Suzuki A,Sakai F,Kobayashi T,Nagahama Y.2006.Knock-down of DMY initiates female pathway in the genetic male medaka,Oryzias latipes.Biochem.Biophys.Res.Commun.,351(4):815-819.
Quitschke W W,Lin Z Y,DePonti-Zilli L,Paterson B M.1989.The beta actin promoter.High levels of transcription depend upon a CCAAT binding factor.J.Biol.Chem.,264(16):9 539-9 546.
Raymond C S,Murphy M W,O'Sullivan M G,Bardwell V J,Zarkower D.2000.Dmrtl,a gene related to worm and fly sexual regulators,is required for mammalian testis differentiation.Genes Dev.,14(20):2 587-2 595.
Raymond C S,Parker E D,Kettlewell J R,Brown L G,Page D C,Kusz K,Jaruzelska J,Reinberg Y,Flej ter W L,Bardwell V J,Hirsch B,Zarkower D.1999.A region of human chromosome 9p required for testis development contains two genes related to known sexual regulators.Hum.Mol.Genet.,8(6):989-996.
Raymond C S,Shamu C E,Shen M M,Seifert K J,Hirsch B,Hodgkin J,Zarkower D.1998.Evidence for evolutionary conservation of sex-determining genes.Nature,391(6668):691-695.
Shao C W,Li Q Y,Chen S L,Zhang P,Lian J M,Hu Q M,Sun B,Jin L J,Liu S S,Wang Z J,Zhao H M,Jin Z H,LiangZ,Li YZ,Zheng Q M,Zhang Y,Wang J,Zhang G J.2014.Epigenetic modification and inheritance in sexual reversal of fish.Genome Res.,24(4):604-615.
Shetty S,Kirby P,Zarkower D,Graves JA M.2002.DMRT1in a ratite bird:evidence for a role in sex determination and discovery of a putative regulatory element.Cytogenet.Genome Res.,99(1-4):245-25 1.
Sinclair A H,Berta P,Palmer M S,Hawkins J R,Griffiths B L,Smith M J,Foster J W,Frischauf A M,Lovell-Badge R,Goodfellow P N.1990.A gene from the human sexdetermining region encodes a protein with homology to a conserved DNA-binding motif.Nature,346(6281):240-244.
Slomiany B A,D'Arigo K L,Kelly M M,Kurtz D T.2000.C/EBPa inhibits cell growth via direct repression of E2FDP-mediated transcription.Mol.Cell.Biol.,20(16):5 986-5 997.
Smith C A,McClive P J,Western P S,Reed K J,Sinclair A H.1999.Conservation of a sex-determining gene.Nature,402(6762):601-602.
Smith C A,Roeszler K N,Ohnesorg T,Cummins D M,Farlie P G,Doran T J,Sinclair A H.2009.The avian Z-linked gene DMRT1 is required for male sex determination in the chicken.Nature,461(7261):267-271.
Sunilkumar G,Connell J P,Smith C W,Reddy A S,Rathore K S.2002.Cotton a-globulin promoter:isolation and functional characterization in transgenic cotton,Arabidopsis,and tobacco.Transgenic Res.,11(4):347-359.
Topilko P,Schneider-Maunoury S,Levi G,Trembleau A,Gourdji D,Driancourt M A,Rao C V,Chamay P.1998.Multiple pituitary and ovarian defects in Krox-24(NGFIA,Egr-1)-targeted mice.Mol Endocrinol.,12(1):107-122.
Tourtellotte W G,Nagarajan R,Auyeung A,Mueller C,Milbrandt J. 1999.Infertility associated with incomplete spermatogenic arrest and oligozoospermia in Egr4-deficient mice.Development,126(22):5 061-5 071.
Tsunoda T,Takagi T.1999.Estimating transcription factor bindability on DNA.Bioinformatics,15(7-8):622-630.
Xia W,Zhou L,Yao B,Li C J,Gui J F.2007.Differential and spermatogenic cell-specific expression of DMRT1 during sex reversal in protogynous hermaphroditic groupers.Mol.Cell.Endocrinol.,263(1-2):1 56-1 72.
Yo shimoto S,Ito M.2011.A ZZ/ZW-type sex determination in Xenopus laevis.FEBS J.,278(7):1 020-1 026.
Zhao L,Svingen T,Ng E T,Koopman P.2015.Female-to-male sex reversal in mice caused by transgenic overexpression of Dmrtl.Development,142(6):1 083-1 088.
Zhou L Q,Yang A G,Liu X Z,Du W,Zhuang Z M.2005.The karyotype of the tonguefish Cynoglossus semilaevis.J.Fish.Sci.China,29(3):417-41 9.(in Chinese with English abstract)