原位杂交和原位PCR技术在鱼类基因定位中的应用

黄梅, 袁仕取, 朱作言

黄梅, 袁仕取, 朱作言. 原位杂交和原位PCR技术在鱼类基因定位中的应用[J]. 水生生物学报, 2001, 25(2): 195-201.
引用本文: 黄梅, 袁仕取, 朱作言. 原位杂交和原位PCR技术在鱼类基因定位中的应用[J]. 水生生物学报, 2001, 25(2): 195-201.
HUANG Mei, YUAN Shi-qu, ZHU Zuo-yan. APPLICATION OF IN SITU HYBRIDIZATION AND IN SITU PCR ON LOCATION OF CENE IN FISHES[J]. ACTA HYDROBIOLOGICA SINICA, 2001, 25(2): 195-201.
Citation: HUANG Mei, YUAN Shi-qu, ZHU Zuo-yan. APPLICATION OF IN SITU HYBRIDIZATION AND IN SITU PCR ON LOCATION OF CENE IN FISHES[J]. ACTA HYDROBIOLOGICA SINICA, 2001, 25(2): 195-201.

原位杂交和原位PCR技术在鱼类基因定位中的应用

基金项目: 

国家自然科学基金(项目编号:39730290)

APPLICATION OF IN SITU HYBRIDIZATION AND IN SITU PCR ON LOCATION OF CENE IN FISHES

  • 摘要: 染色体原位杂交(in situ hybridization, ISH)是基因物理定位的主要方法之一,它根据核酸分子碱基互补配对的原理,将标记的外源核酸探针与染色体上变性处理后的单链DNA互补配对,再经检测而将靶顺序在染色体上的位置显示出来.原位PCR技术是将原位杂交与PCR技术有机的结合,在原位扩增目的DNA片段,并在原位检测其扩增产物,因而兼有PCR及原位杂交技术二者的优越性:既可以同时获得组织及细胞的形态结构信息与分子信息,进行定性、定位分析,又具有比原位杂交更好的敏感性与专一性. 染色体原位杂交和原位PCR这两种技术是基因定位中简便而直接的方法,随着基因组研究和转基因生物研究的深入与与发展,阐明各种基因在染色体上的具体位置愈加重要.本文拟从原位杂交和原位PCR的发展、原理、方法和在鱼类基因的染色体定位等方面进行评述和讨论.
  • [1] Gall J,G,Pardue H L,Formation and detection of RNA-DNA hybrid molecules in cytoligical preparations [J].Pro.Natl.Acad Sci U.S.A,1969,63:378—381[2] Gerhard D S,Kawasaki E.S,Bancroft F.C,et al,Localization of a unique gene by direct hybridization in situ [J].Pro.Natl.Acad Sci U.S.A,1981.78:3755—3759[3] Harper M E,Sarunders G F.Localization of the human insulin gene to the distal end of the short arm of chromosome 11 [J].Pro.Natl.Acad Sci.U.S.A,1981 78:4458—4460[4] Langer-Safer P R Levine M,Ward D C.Immuunological method for mapping genes on drosophila polytene chromosomes[J].Pro.Natl.Acad Sci U.S.A,1982,79:4381—4385[5] Cannizzaro L A,Emanuel B S.In situ hybridization and translocation breakpoint mapping.[J] Cytogenet.Cell.Genet.1985,39:179—183[6] Bostock C J,Clark E M.Satellite DNA in large marker chromosomes of methotrecate-resistant mouse cells.[J] Cell,1980,19:709—715[7] John H.A,Singh L.RNA-DNA hybrids at the cytological level[J] Nature,1969,223:582—587[8] Pendas A.M,Moran P,Eva Garcia-Vazquez.Orgenization and chromosomal location of the major histone cluster in Brown Trout,Atlantic Salmon and Rainbow Trout.[J] Chromosoma,1994 103:147—152[9] Reed K M,Phillips R B Molecular characterization and cytogenetic analysis of highly reoeated DNAs of Lake Trout,Salvelinus Namaycush.[J].Chromosoma,1995,104:242—251[10] Reed K M,Phillips R B.Molecular cytogenetic analysis of the double-cma3 chromosome of Lake Trout,Salvelins Namaycush [J].Cytogenet.Cell Genet.1995,70:104—107[11] 张锡元,童素红,将建桥.草鱼的bHCG基因同类物及其染色体定位[J].遗传学报,1989,16(4):299—304[12] 李 奎,余其兴,毛 勇,等.用地高辛标记原位杂交技术定位黄鳝rRNA基因于二价染色体3q12-q24和7q14-q26 [J].遗传学报,1995,22(2):97—102[13] 王泽群,朱作言.鲤鱼基因组DNA重复序列CR1的染色体原位杂交定位[J].科学通报,1999,44(20):2182—2186[14] 李书鸿,郭绍东,孙方臻.用整体原位杂交法研究微管蛋白基因在斑马鱼胚胎中的表达[J].科学通报,1998,43(17):1855—1858[15] Chollet A,Kawashima E M.Biotin labelled synthetic oligodeoxy ribonucleotides synthesis and uses as hybridization probes [J].Nuclear Acid Research.,1985,13:1529—1541[16] Garrido-Ramos M A,Kawasaki E S,Bancroft F C.Cloning and Characterization of a fish centromeric satellite DNA [J].Cytogenet,Cell Genet.,1994,65:233—237[17] Pendas A M,Moran P,Eva Garcia-Vazque.Chromosomal mapping and nucleotide sequence of two tandem repeats of Atlantic salmon 5s-Rdna [J].Cytogenet.Cell Genet.,1994,64:31—36[18] Smith V T H B M,Rose M R.Combined GTG-banding and nonradioactive in situ hybridization improves characterization of complex kiaryotypes [J].Cytogenet.Cell Genet.,1990,54:20—23[19] Speleman F,Ihssen P E,Pieper F R.Characterization of a de novo duplication of 11p14-p13,using fluorescent in situ hybridization and southern hybridization [J].Cytogenet.Cell Genet.1991,56:129—131[20] Phillips R B,Reed K M.Application of flourescence in situ hybridization(FISH)techniques to fish genetics:a review[J].Aquaculture,1996,140:197—216[21] Koch J E,Kolvraa S,Peetersssen K B,et al Oligonnucleotide-priming nethods for the chromosome-specific labeling of alpha satellite DNA in situ [J],Chromosoma,1989,98:259—265[22] Mullis K,Faloona F,Scharf S,et al.Specific enzymatic amplification of DNA in vitro the polymerase chain reaction [J].Cold Spring Harber Symp Quant Biol.1986,51:263—273[23] 杜丽萍,余其兴,郭一清.黄鳝二价上Px基因外显特异引物原位DNA合成[J] 遗传学报,1998,(25)(1):28—33[24] Troyer D L,Goad D W,Xie H,et al.Use of direction in situ single-copy(DISC)PCR to physically map five porcine microsatellites [J].Cytogenet Cell Genet 1994,67:199—204[25] 余先觉.中国淡水鱼类染色体[M].北京:科学出版社,1989[26] Zhang X Y,Jiang H B,Li L J.In situ amplification of DNA fragments specific for human Y chromosome in cellular nuclei by PCR [J].Science In China(Series C)1996,39(1):45—52[27] Nuovo G J,Macconnell P,Forde A.et al.Detection of human papillomavirus DNA in formalin-fixed tissues by in situ hybridization after amplification by polymerase chain reaction[J].Am.J.Pathol.1991,139:847—854[28] Long A.A,Komminoth P.Lee E.Comparison of indirect and direct in situ polymerase chain reaction in cell preparations and tissue sections[J].Histochemistry 99:151—162[29] Nuovo G J,Lidonnici K,Mac Connell P,et al.Intracellular localization of polymerase chain reaction(PCR)-amplified hepatitis C cDNA[J].Am J Surg Pathol.1993,17(7):683—690[30] 朱作言,许克圣,李国华,等 转基因鱼模型的建立[J].中国科学,1989(B)2:147—155[31] Zhu Z,Li G,He L,et al.Novel gene transfer into the fertilized eggs of goldfish(Carassius auratus L.1758)[J].Z angew Ichthyol,1985,1:31—34

    Gall J,G,Pardue H L,Formation and detection of RNA-DNA hybrid molecules in cytoligical preparations [J].Pro.Natl.Acad Sci U.S.A,1969,63:378—381[2] Gerhard D S,Kawasaki E.S,Bancroft F.C,et al,Localization of a unique gene by direct hybridization in situ [J].Pro.Natl.Acad Sci U.S.A,1981.78:3755—3759[3] Harper M E,Sarunders G F.Localization of the human insulin gene to the distal end of the short arm of chromosome 11 [J].Pro.Natl.Acad Sci.U.S.A,1981 78:4458—4460[4] Langer-Safer P R Levine M,Ward D C.Immuunological method for mapping genes on drosophila polytene chromosomes[J].Pro.Natl.Acad Sci U.S.A,1982,79:4381—4385[5] Cannizzaro L A,Emanuel B S.In situ hybridization and translocation breakpoint mapping.[J] Cytogenet.Cell.Genet.1985,39:179—183[6] Bostock C J,Clark E M.Satellite DNA in large marker chromosomes of methotrecate-resistant mouse cells.[J] Cell,1980,19:709—715[7] John H.A,Singh L.RNA-DNA hybrids at the cytological level[J] Nature,1969,223:582—587[8] Pendas A.M,Moran P,Eva Garcia-Vazquez.Orgenization and chromosomal location of the major histone cluster in Brown Trout,Atlantic Salmon and Rainbow Trout.[J] Chromosoma,1994 103:147—152[9] Reed K M,Phillips R B Molecular characterization and cytogenetic analysis of highly reoeated DNAs of Lake Trout,Salvelinus Namaycush.[J].Chromosoma,1995,104:242—251[10] Reed K M,Phillips R B.Molecular cytogenetic analysis of the double-cma3 chromosome of Lake Trout,Salvelins Namaycush [J].Cytogenet.Cell Genet.1995,70:104—107[11] 张锡元,童素红,将建桥.草鱼的bHCG基因同类物及其染色体定位[J].遗传学报,1989,16(4):299—304[12] 李 奎,余其兴,毛 勇,等.用地高辛标记原位杂交技术定位黄鳝rRNA基因于二价染色体3q12-q24和7q14-q26 [J].遗传学报,1995,22(2):97—102[13] 王泽群,朱作言.鲤鱼基因组DNA重复序列CR1的染色体原位杂交定位[J].科学通报,1999,44(20):2182—2186[14] 李书鸿,郭绍东,孙方臻.用整体原位杂交法研究微管蛋白基因在斑马鱼胚胎中的表达[J].科学通报,1998,43(17):1855—1858[15] Chollet A,Kawashima E M.Biotin labelled synthetic oligodeoxy ribonucleotides synthesis and uses as hybridization probes [J].Nuclear Acid Research.,1985,13:1529—1541[16] Garrido-Ramos M A,Kawasaki E S,Bancroft F C.Cloning and Characterization of a fish centromeric satellite DNA [J].Cytogenet,Cell Genet.,1994,65:233—237[17] Pendas A M,Moran P,Eva Garcia-Vazque.Chromosomal mapping and nucleotide sequence of two tandem repeats of Atlantic salmon 5s-Rdna [J].Cytogenet.Cell Genet.,1994,64:31—36[18] Smith V T H B M,Rose M R.Combined GTG-banding and nonradioactive in situ hybridization improves characterization of complex kiaryotypes [J].Cytogenet.Cell Genet.,1990,54:20—23[19] Speleman F,Ihssen P E,Pieper F R.Characterization of a de novo duplication of 11p14-p13,using fluorescent in situ hybridization and southern hybridization [J].Cytogenet.Cell Genet.1991,56:129—131[20] Phillips R B,Reed K M.Application of flourescence in situ hybridization(FISH)techniques to fish genetics:a review[J].Aquaculture,1996,140:197—216[21] Koch J E,Kolvraa S,Peetersssen K B,et al Oligonnucleotide-priming nethods for the chromosome-specific labeling of alpha satellite DNA in situ [J],Chromosoma,1989,98:259—265[22] Mullis K,Faloona F,Scharf S,et al.Specific enzymatic amplification of DNA in vitro the polymerase chain reaction [J].Cold Spring Harber Symp Quant Biol.1986,51:263—273[23] 杜丽萍,余其兴,郭一清.黄鳝二价上Px基因外显特异引物原位DNA合成[J] 遗传学报,1998,(25)(1):28—33[24] Troyer D L,Goad D W,Xie H,et al.Use of direction in situ single-copy(DISC)PCR to physically map five porcine microsatellites [J].Cytogenet Cell Genet 1994,67:199—204[25] 余先觉.中国淡水鱼类染色体[M].北京:科学出版社,1989[26] Zhang X Y,Jiang H B,Li L J.In situ amplification of DNA fragments specific for human Y chromosome in cellular nuclei by PCR [J].Science In China(Series C)1996,39(1):45—52[27] Nuovo G J,Macconnell P,Forde A.et al.Detection of human papillomavirus DNA in formalin-fixed tissues by in situ hybridization after amplification by polymerase chain reaction[J].Am.J.Pathol.1991,139:847—854[28] Long A.A,Komminoth P.Lee E.Comparison of indirect and direct in situ polymerase chain reaction in cell preparations and tissue sections[J].Histochemistry 99:151—162[29] Nuovo G J,Lidonnici K,Mac Connell P,et al.Intracellular localization of polymerase chain reaction(PCR)-amplified hepatitis C cDNA[J].Am J Surg Pathol.1993,17(7):683—690[30] 朱作言,许克圣,李国华,等 转基因鱼模型的建立[J].中国科学,1989(B)2:147—155[31] Zhu Z,Li G,He L,et al.Novel gene transfer into the fertilized eggs of goldfish(Carassius auratus L.1758)[J].Z angew Ichthyol,1985,1:31—34

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  • 收稿日期:  2000-01-28
  • 修回日期:  2000-04-16
  • 发布日期:  2001-03-24

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