微卫星DNA多态性分析在常用近交系小鼠遗传监测中的应用研究
|
摘要
近交系小鼠是医学生物学研究中广泛应用的实验动物，其本身的质量对实验结果的可比性、可重复性和准确性起着决定性作用。随着生物医学的发展，对实验动物质量的要求越来越高，因而对实验动物质量的遗传监测方法也提出了更高的要求。微卫星DNA标记与目前国标推荐的生化标记检测方法相比，具有敏感性高、样本需要量少、简便快速等优点，在遗传多样性研究领域有着广阔的应用前景。目前，国内应用微卫星DNA标记分析只是对其可行性进行了尝试，微卫星的选取多是随机的，因而多态性较差，不能对近交系进行有效监测，尚未建立针对我国常用近交系小鼠进行系统而有效遗传检测的微卫星位点组合(microsatellites panel)，微卫星DNA遗传检测的方法及判定的标准上在国内尝属空白，因此本研究对于提高我国实验动物遗传监测技术水平，促进我国实验动物科学的发展，有着重要的现实意义。
本实验从发达国家已成熟应用并有着较好基因频率分布的微卫星中，筛选了位于不同染色体上的16个微卫星位点，应用聚合酶链反应(PCR)、聚丙酰胺凝胶电泳、银染染色和凝胶成像分析等技术，对国内常用的10种近交系小鼠进行了微卫星DNA多态性分析，建立了应用微卫星DNA进行小鼠近交系遗传监测的方法和可供参考的微卫星DNA多态性图谱。实验表明其中14个位点能特异地反映出10个近交系小鼠的品系特性，可作为近交系小鼠遗传监测的微卫星位点，用于小鼠遗传质量常规检测和遗传背景分析。并用这14个微卫星位点对全国7个地区的11个BALB／c小鼠群进行遗传监测，检测出了生化标记所不能检出的遗传变异，说明应用微卫星DNA标记对近交系小鼠进行遗传监测，方法快速、简便、敏感。
首先，本研究通过对PCR反应液组成和热循环条件的优化，建立了PCR扩增微卫星DNA的方法。运用此方法，微卫星DNA可稳定、特异、有效地扩增。确立了针对每一位点的最佳M g~(2+)浓度和最佳退火温度。
实验所选用的我国10个常用近交系小鼠是BALB／c、C57BL／6J、AMMS／1、C3H／He、DBA／2N、129／Sv、FVB／N、615、TA_1、TA_2，应用筛选的16对微卫星引物(位点名称为：D1Mit365、D2Mit30、D3Mit51、D4Mit235、D5Mit48、D6Mit102、D7Mit281、D8Mit113、D9Mit23、D10Mit180、D11Mit128、D12Mit147、D13 Mit88、D14Mit102、D15Mit5、D17Mit36)
中文摘要
进行了DNA多态性分析，并与目前常规应用的生化标记分析法进行了比较。结果表明:
除了DSMi t 1 13、Dl 3 Mit88没能有效扩增外，其余14个位点均有稳定扩增效果，10个
品系的近交系小鼠在这14个位点上均呈现一条清晰图带，且在同一品系内图带泳动距
离一致，表明所检测的小鼠在研究位点上表现为纯合，遗传背景均一，没有发生遗传变
异，这与生化位点检测结果一致。在这14个位点，10个品系小鼠间呈现显著多态性，
并能特异地反映出10个近交系小鼠的品系特性。说明该方法可通过这14个微卫星位点
进行遗传监测。通过不同品系间相似系数的比较，可进行品系间亲缘关系远近的判断。
品系间相似系数范围在14.3%( C57与129)一92.90&(TAI与TAZ)之间，TA、与TAZ仅在
DgMit23这一个位点有差异，相似系数高达92.9ry0，而129/Sv与其余品系间相似系数远
远低于其它品系间的相似系数，说明TA、与TA:有着较近的亲缘关系，129/sv与其余品
系间亲缘关系较远。
其次，采用所筛选的14个微卫星位点对北京、上海、沈阳、哈尔滨、广州、重庆、
长春等n家小鼠生产厂家所提供的BALB/c小鼠进行遗传监测，结果表明:北京、上
海、哈尔滨等7家小鼠生产厂家所提供的BALB/c小鼠在同一群体内表型一致，且在各
位点不同群体间也无差异，表明所检测的小鼠在14个位点上表现为纯合，未发生遗传变
异。沈阳、广州、重庆、长春4家的BALB/C小鼠的8个位点在群体内表型不一致，其
中重庆地区小鼠在7个位点(DIMit365、DZMit3O、D3Mit51、DSMit48、D6Mi七102、D7Mit281、
DIOMit18O)发生遗传污染或遗传变异，广州地区一单位(穗1)小鼠在4个位点(DZMit30、
DSMit48、D6MitlOZ、DIOMit18O)发生变异，沈阳地区小鼠在位点D15mits发生变异，
长春地区小鼠在位点DZMit30存在变异。这些变异表现为二种情况的多态性:(1)电泳
虽只表现为一条带，但彼此间泳动距离不相一致:(2)在排除影子带后仍表现两条图带;
这二种情况的多态性的出现都表明群体在这8个位点上发生了遗传变异，而生化位点检
测没能检测出这些变异，说明微卫星DNA标记要比生化位点标记更准确、有效。本实验
也为了解我国各地区近交系小鼠遗传概貌，规范引种、供种，实现种质保证提供理论依
据。
总之，本实验中的14个位点能特异地反映出近交系小鼠的品系特性，可作为近交
系小鼠遗传监测的微卫星位点，用于常规检测小鼠遗传质量和遗传背景分析等。
The inbred mouse is the most widely studied animal model in major disciplines of biomedical research. The quality of these animals is a decisive influence on comparative, repetition and veracity of the experiment. With the develop of biomedical research, the more require of qualified mice, the more efficacious genetic monitoring will be needed. Now, methods used in genetic quality control involve the use or biochemical markers, immunologic methods, strain-specific typing sera, morphologic characteristics such as the shape of the mandible, and skin grafting. Since 1980, DNA marker came forth one after another. Among them microsatellite DNA possesses various, scattered wide, high degree of polymorphism, easy to communicate and so on . So this approach is the most usefulness, and it become an ideal genetic marker for gene localization, gene maping, population study and individual identificatinon.
Microsatellites consist of around 10-50copies of motifs from 1 to 6 that can occur in perfect tandem repetition. It occurs frequently and randomly in all eukaryotic DNAs examined. Microsatellite DNA are ideal genetic markers due to the high degree of polymorphism and the fact that the variation is readily analyzed using the polymerase chain reaction (PCR).
The aim of genetic monitoring is to checking the variance of the animal have been done or not, whether interfuse other strains, which insures that the strains according with the require of colony . At present, the methods based on allozyme biochemistry are the National Standard instructed. Methods that using microsatellite DNA would be more useful for genetic monitoring than methods based on allozyme biochemistry because the genome itself is being tested rather than a protein product and a larger portion of the genome can be sampled, and easy to distinguish. Methods that using microsatellite DNA had abundant microsatellite
loci (over 7300, before 1999) can be identified. Applying enough microsatellite loci will present abundant straps and well polymorphism, which can reflection inherit and variation of roundly genome. In addition, the PCR-based microsatellite analysis is a fast and economical way for detecting genetic contamination . So we should select microsatellite DNA which is polymorphisms as genetic monitoring markers to determining the strains' origin and genetic background of inbred mice. Now only feasibility has been report, and no reports on standards and practicality has been found in our country.
With the optimization of components of reaction buffer and amplification parameter, PCR for amplification microsatellite DNA was finally set up. Using the techniques, microsatellite DNA can amplified efficaciously. The final concentrations of Mg2+ was 1 .5 - 3. 0 mmol/L, annealing temperature was 50℃-65℃.The condition for the PCR amplify were ,94℃for 3min, SOcycles of 94℃ for 30s, 50~65"C for 30s, 72℃ for Imin, finally at 72℃for Imin, then store at 4℃. Ten kinds of inbred strain mice including C57BL / 6J, C3H/He, TA,, TA2, 615, BALB/c ,DBA/2N,129/Sv,FVB/N, AMMS/1 were investigated by PCR analysis. This is the first time to genetic monitoring on commonly used inbred mouse strains by the 14 microsatellite DNA . It showed that all these microsatellites DNA loci display single allelic gene band. Fourteen loci are polymorphisms between the inbred strains . The polymorphisms of DlMit365, D2Mit30, D3Mit51, D5Mit48, D6Mit102, D10Mit180, DllMit128, D12Mit147, D14Mitl02 and D17MU36 are significant .These results sugge
st that these mice tested meet the request of inbred strain. The genetic background of TA, was similar with that of TA2, the similarity indices were from 14. 3%( C57 between 129)to 92. 9%( TA, between TA2).This means that TA, and TA2 had closer genetic relationship, C57 and 129 had farther relationship. Strain 129/Sv is clearly placed at the most deeply diverged of the domesticus strains represented. So it is possible to discriminate between any pair of strains,even closely related and congenic strains. Screened loci showing marked polymorphisms typically
本实验从发达国家已成熟应用并有着较好基因频率分布的微卫星中，筛选了位于不同染色体上的16个微卫星位点，应用聚合酶链反应(PCR)、聚丙酰胺凝胶电泳、银染染色和凝胶成像分析等技术，对国内常用的10种近交系小鼠进行了微卫星DNA多态性分析，建立了应用微卫星DNA进行小鼠近交系遗传监测的方法和可供参考的微卫星DNA多态性图谱。实验表明其中14个位点能特异地反映出10个近交系小鼠的品系特性，可作为近交系小鼠遗传监测的微卫星位点，用于小鼠遗传质量常规检测和遗传背景分析。并用这14个微卫星位点对全国7个地区的11个BALB／c小鼠群进行遗传监测，检测出了生化标记所不能检出的遗传变异，说明应用微卫星DNA标记对近交系小鼠进行遗传监测，方法快速、简便、敏感。
首先，本研究通过对PCR反应液组成和热循环条件的优化，建立了PCR扩增微卫星DNA的方法。运用此方法，微卫星DNA可稳定、特异、有效地扩增。确立了针对每一位点的最佳M g~(2+)浓度和最佳退火温度。
实验所选用的我国10个常用近交系小鼠是BALB／c、C57BL／6J、AMMS／1、C3H／He、DBA／2N、129／Sv、FVB／N、615、TA_1、TA_2，应用筛选的16对微卫星引物(位点名称为：D1Mit365、D2Mit30、D3Mit51、D4Mit235、D5Mit48、D6Mit102、D7Mit281、D8Mit113、D9Mit23、D10Mit180、D11Mit128、D12Mit147、D13 Mit88、D14Mit102、D15Mit5、D17Mit36)
中文摘要
进行了DNA多态性分析，并与目前常规应用的生化标记分析法进行了比较。结果表明:
除了DSMi t 1 13、Dl 3 Mit88没能有效扩增外，其余14个位点均有稳定扩增效果，10个
品系的近交系小鼠在这14个位点上均呈现一条清晰图带，且在同一品系内图带泳动距
离一致，表明所检测的小鼠在研究位点上表现为纯合，遗传背景均一，没有发生遗传变
异，这与生化位点检测结果一致。在这14个位点，10个品系小鼠间呈现显著多态性，
并能特异地反映出10个近交系小鼠的品系特性。说明该方法可通过这14个微卫星位点
进行遗传监测。通过不同品系间相似系数的比较，可进行品系间亲缘关系远近的判断。
品系间相似系数范围在14.3%( C57与129)一92.90&(TAI与TAZ)之间，TA、与TAZ仅在
DgMit23这一个位点有差异，相似系数高达92.9ry0，而129/Sv与其余品系间相似系数远
远低于其它品系间的相似系数，说明TA、与TA:有着较近的亲缘关系，129/sv与其余品
系间亲缘关系较远。
其次，采用所筛选的14个微卫星位点对北京、上海、沈阳、哈尔滨、广州、重庆、
长春等n家小鼠生产厂家所提供的BALB/c小鼠进行遗传监测，结果表明:北京、上
海、哈尔滨等7家小鼠生产厂家所提供的BALB/c小鼠在同一群体内表型一致，且在各
位点不同群体间也无差异，表明所检测的小鼠在14个位点上表现为纯合，未发生遗传变
异。沈阳、广州、重庆、长春4家的BALB/C小鼠的8个位点在群体内表型不一致，其
中重庆地区小鼠在7个位点(DIMit365、DZMit3O、D3Mit51、DSMit48、D6Mi七102、D7Mit281、
DIOMit18O)发生遗传污染或遗传变异，广州地区一单位(穗1)小鼠在4个位点(DZMit30、
DSMit48、D6MitlOZ、DIOMit18O)发生变异，沈阳地区小鼠在位点D15mits发生变异，
长春地区小鼠在位点DZMit30存在变异。这些变异表现为二种情况的多态性:(1)电泳
虽只表现为一条带，但彼此间泳动距离不相一致:(2)在排除影子带后仍表现两条图带;
这二种情况的多态性的出现都表明群体在这8个位点上发生了遗传变异，而生化位点检
测没能检测出这些变异，说明微卫星DNA标记要比生化位点标记更准确、有效。本实验
也为了解我国各地区近交系小鼠遗传概貌，规范引种、供种，实现种质保证提供理论依
据。
总之，本实验中的14个位点能特异地反映出近交系小鼠的品系特性，可作为近交
系小鼠遗传监测的微卫星位点，用于常规检测小鼠遗传质量和遗传背景分析等。
The inbred mouse is the most widely studied animal model in major disciplines of biomedical research. The quality of these animals is a decisive influence on comparative, repetition and veracity of the experiment. With the develop of biomedical research, the more require of qualified mice, the more efficacious genetic monitoring will be needed. Now, methods used in genetic quality control involve the use or biochemical markers, immunologic methods, strain-specific typing sera, morphologic characteristics such as the shape of the mandible, and skin grafting. Since 1980, DNA marker came forth one after another. Among them microsatellite DNA possesses various, scattered wide, high degree of polymorphism, easy to communicate and so on . So this approach is the most usefulness, and it become an ideal genetic marker for gene localization, gene maping, population study and individual identificatinon.
Microsatellites consist of around 10-50copies of motifs from 1 to 6 that can occur in perfect tandem repetition. It occurs frequently and randomly in all eukaryotic DNAs examined. Microsatellite DNA are ideal genetic markers due to the high degree of polymorphism and the fact that the variation is readily analyzed using the polymerase chain reaction (PCR).
The aim of genetic monitoring is to checking the variance of the animal have been done or not, whether interfuse other strains, which insures that the strains according with the require of colony . At present, the methods based on allozyme biochemistry are the National Standard instructed. Methods that using microsatellite DNA would be more useful for genetic monitoring than methods based on allozyme biochemistry because the genome itself is being tested rather than a protein product and a larger portion of the genome can be sampled, and easy to distinguish. Methods that using microsatellite DNA had abundant microsatellite
loci (over 7300, before 1999) can be identified. Applying enough microsatellite loci will present abundant straps and well polymorphism, which can reflection inherit and variation of roundly genome. In addition, the PCR-based microsatellite analysis is a fast and economical way for detecting genetic contamination . So we should select microsatellite DNA which is polymorphisms as genetic monitoring markers to determining the strains' origin and genetic background of inbred mice. Now only feasibility has been report, and no reports on standards and practicality has been found in our country.
With the optimization of components of reaction buffer and amplification parameter, PCR for amplification microsatellite DNA was finally set up. Using the techniques, microsatellite DNA can amplified efficaciously. The final concentrations of Mg2+ was 1 .5 - 3. 0 mmol/L, annealing temperature was 50℃-65℃.The condition for the PCR amplify were ,94℃for 3min, SOcycles of 94℃ for 30s, 50~65"C for 30s, 72℃ for Imin, finally at 72℃for Imin, then store at 4℃. Ten kinds of inbred strain mice including C57BL / 6J, C3H/He, TA,, TA2, 615, BALB/c ,DBA/2N,129/Sv,FVB/N, AMMS/1 were investigated by PCR analysis. This is the first time to genetic monitoring on commonly used inbred mouse strains by the 14 microsatellite DNA . It showed that all these microsatellites DNA loci display single allelic gene band. Fourteen loci are polymorphisms between the inbred strains . The polymorphisms of DlMit365, D2Mit30, D3Mit51, D5Mit48, D6Mit102, D10Mit180, DllMit128, D12Mit147, D14Mitl02 and D17MU36 are significant .These results sugge
st that these mice tested meet the request of inbred strain. The genetic background of TA, was similar with that of TA2, the similarity indices were from 14. 3%( C57 between 129)to 92. 9%( TA, between TA2).This means that TA, and TA2 had closer genetic relationship, C57 and 129 had farther relationship. Strain 129/Sv is clearly placed at the most deeply diverged of the domesticus strains represented. So it is possible to discriminate between any pair of strains,even closely related and congenic strains. Screened loci showing marked polymorphisms typically
引文
[1] Tautz D. Hypervariability of simple sequences as a general source for polymorphic DNA markers [J].Nucleic Acids Res, 1989,17(16):6463～6467
[2] Hsmsda H, Petrino M G, Kskunaga T.A novel repeated element with Z-DNA-forming potential is widely found in evolutionarily diverse eukaryotic genomes [J].Proc Natl Acad Sci, 1982,79(21):6465～6469
[3] Peters J, Selley R L, Cocking Y. Mouse DNA segment[J].Mouse Genome. 1996,94(2):404～473
[4] Dietrich W, Katz H, Lincoln S E et al.A genetic map of the mouse suitable for typing intraspecific crosses. Genetics, 1992,131:423～447
[5] Thomas E. Le Voyer, KENt W. Hunter. Microsatellite DNA variants among the FVB/NJ, C58/J, and I/LnJ mouse strains[J].Mamm Genome, 1999,10,542～543
[6] J.H. Slingsby, M.B. Hogarth et al. New microsatellite polymorphisms identified between C57BL/6, C57BL/10, and C57BL/KsJ inbred mouse strains[J], immunogenetics, 1995,43:72-75
[7] F. Benavides, E. Glasscock, et al. PCR-based microsatellite analysis for differentiation and genetic monitoring of nine inbred SENCAR mouse strains[J].Laboratory Amimals, 2001,35:157-162
[8] 孟雁，宋昌祜，奥本正昭，等．10种近交系小鼠的微卫星位点分析及其在遗传监测中应用的探讨(一)[J]．中国实验动物学报，1996，4(1)：5-9
[9] 曾苹，张连峰，程振华，等．利用PCR技术分析微卫星DNA进行小鼠的遗传监测[J]．中国实验动物学杂志，1995，5(4)：225-228
[10] 张树辉，魏泓，史景泉．近交系小鼠微卫星DNA多态性的研究[J]．遗传，2000，22(6)：375～378
[11] 谢建云，邵伟娟，高诚．近交系小鼠微卫星位点遗传检测方法的建立[J]．上海交通大学学报(农业科学版)2002，20(2)：100-104
[12] Mouse Genome Database(MGD),Mouse Genome Informatics Project, The
Jakson Laboratory, Bar Harbor, ME. World Wide Web(URL:http// www. informatics, jax. org)
[13] Murray V, Chutima M, England PR. The determination of the sequences present in the shadow bands of a dinucleiotide repeat by PCR[J]. Nucleic Acids Res, 1993,21(10): 2395-2398
[14] Hauge XY, Litt M.A study of the origin of "shadow band" seen when typing dinucleiotide repeat polymorphisms by the PCR[J]. Hum Mol Genet,,1993,2(4): 411-415
[15] 武辉，张思仲，肖翠英．三核苷酸重复序列PCR扩增中影子带的产生及其消除的方法[J]．中华医学遗传学杂志，1998，15(1)：42-45
[16] 赵红珊，黄尚志，等．短重复序列PCR产物变性胶电泳影子带新解——正负链电泳行为的差异[J]．中华医学遗传杂志，1999，16(5)：328-330
[17] Edwards AL, Civitello A, Holly A, et al. DNA typing and genetics mapping with trimeric and tetrametic tandem trpeats[J].Am J Hum Genet, 1991,49:746-756
[18] 高玉彤，来茂德．银染色法检测微卫星不稳定性[J]．临床与实验病理学杂志，1998，14：193-194
[19] Roxanne Y. Walder, MICHAEL R, et al. Shot tandem repeat polymorphic markers for the rat genome from marker-selected libraries[J].Mammalian Genome, 1998,9:1013-1021
[20] 陈振文，李瑞生，等．DNA指纹图与生化标记分析对近交系小鼠遗传临测的比较[J]．中国兽医学报，2001，21(6)：627-630
[22] Leonard C. Schalkwyk, Msrtin Jung, et al. Panel of microsatellite markers for whole-genome scans and radiation hybrid mapping and a mouse family tree[J].Genome Research, 1999,9:878-887
[23] 吴梅筠．法医物证学[M]．北京：人民军医出版社，1998，200
[24] 盛岩，郑蔚虹，等．微卫星标记在种群生物学研究中的应用[J]．植物生态学报，2002，26(增刊)：119-126
[25] James F. Willott, Jeremy G. Turner, et al. The BALB/cmouse as an animal model for progressive sensorineural hearing loss [J].Hearing
Research, 1998,115:162-174
[26] 李厚达．实验动物学[M]．北京：中国农业出版社，2003，247
[27] 孙伟．微卫星DNA的研究与应用[J]．黑龙江畜牧兽医，2001，(3)10—12．
[28] 李牧尧．两种新的DNA多态标记—小卫星和微卫星DNA[J]．国外医学-遗传学分册，1992，15(4)，172-173
[29] 李瑞生，陈振文，等．近交系大鼠微卫星DNA与生化标记分析的比较研究[J]．中国实验动物学杂志，2002，12(4)：225-228
[30] Brant J. Bassam, Gustavo Caetano-Anollés, et al. Fast and sensitive silver staining of DNA in polyacrylamide gels[J].Analytical biochemistry, 1991, 196:80-83
[31] 张艳，张树义．微卫星方法简介[J]，动物学杂志，1999，34(2)：42-45
[2] Hsmsda H, Petrino M G, Kskunaga T.A novel repeated element with Z-DNA-forming potential is widely found in evolutionarily diverse eukaryotic genomes [J].Proc Natl Acad Sci, 1982,79(21):6465～6469
[3] Peters J, Selley R L, Cocking Y. Mouse DNA segment[J].Mouse Genome. 1996,94(2):404～473
[4] Dietrich W, Katz H, Lincoln S E et al.A genetic map of the mouse suitable for typing intraspecific crosses. Genetics, 1992,131:423～447
[5] Thomas E. Le Voyer, KENt W. Hunter. Microsatellite DNA variants among the FVB/NJ, C58/J, and I/LnJ mouse strains[J].Mamm Genome, 1999,10,542～543
[6] J.H. Slingsby, M.B. Hogarth et al. New microsatellite polymorphisms identified between C57BL/6, C57BL/10, and C57BL/KsJ inbred mouse strains[J], immunogenetics, 1995,43:72-75
[7] F. Benavides, E. Glasscock, et al. PCR-based microsatellite analysis for differentiation and genetic monitoring of nine inbred SENCAR mouse strains[J].Laboratory Amimals, 2001,35:157-162
[8] 孟雁，宋昌祜，奥本正昭，等．10种近交系小鼠的微卫星位点分析及其在遗传监测中应用的探讨(一)[J]．中国实验动物学报，1996，4(1)：5-9
[9] 曾苹，张连峰，程振华，等．利用PCR技术分析微卫星DNA进行小鼠的遗传监测[J]．中国实验动物学杂志，1995，5(4)：225-228
[10] 张树辉，魏泓，史景泉．近交系小鼠微卫星DNA多态性的研究[J]．遗传，2000，22(6)：375～378
[11] 谢建云，邵伟娟，高诚．近交系小鼠微卫星位点遗传检测方法的建立[J]．上海交通大学学报(农业科学版)2002，20(2)：100-104
[12] Mouse Genome Database(MGD),Mouse Genome Informatics Project, The
Jakson Laboratory, Bar Harbor, ME. World Wide Web(URL:http// www. informatics, jax. org)
[13] Murray V, Chutima M, England PR. The determination of the sequences present in the shadow bands of a dinucleiotide repeat by PCR[J]. Nucleic Acids Res, 1993,21(10): 2395-2398
[14] Hauge XY, Litt M.A study of the origin of "shadow band" seen when typing dinucleiotide repeat polymorphisms by the PCR[J]. Hum Mol Genet,,1993,2(4): 411-415
[15] 武辉，张思仲，肖翠英．三核苷酸重复序列PCR扩增中影子带的产生及其消除的方法[J]．中华医学遗传学杂志，1998，15(1)：42-45
[16] 赵红珊，黄尚志，等．短重复序列PCR产物变性胶电泳影子带新解——正负链电泳行为的差异[J]．中华医学遗传杂志，1999，16(5)：328-330
[17] Edwards AL, Civitello A, Holly A, et al. DNA typing and genetics mapping with trimeric and tetrametic tandem trpeats[J].Am J Hum Genet, 1991,49:746-756
[18] 高玉彤，来茂德．银染色法检测微卫星不稳定性[J]．临床与实验病理学杂志，1998，14：193-194
[19] Roxanne Y. Walder, MICHAEL R, et al. Shot tandem repeat polymorphic markers for the rat genome from marker-selected libraries[J].Mammalian Genome, 1998,9:1013-1021
[20] 陈振文，李瑞生，等．DNA指纹图与生化标记分析对近交系小鼠遗传临测的比较[J]．中国兽医学报，2001，21(6)：627-630
[22] Leonard C. Schalkwyk, Msrtin Jung, et al. Panel of microsatellite markers for whole-genome scans and radiation hybrid mapping and a mouse family tree[J].Genome Research, 1999,9:878-887
[23] 吴梅筠．法医物证学[M]．北京：人民军医出版社，1998，200
[24] 盛岩，郑蔚虹，等．微卫星标记在种群生物学研究中的应用[J]．植物生态学报，2002，26(增刊)：119-126
[25] James F. Willott, Jeremy G. Turner, et al. The BALB/cmouse as an animal model for progressive sensorineural hearing loss [J].Hearing
Research, 1998,115:162-174
[26] 李厚达．实验动物学[M]．北京：中国农业出版社，2003，247
[27] 孙伟．微卫星DNA的研究与应用[J]．黑龙江畜牧兽医，2001，(3)10—12．
[28] 李牧尧．两种新的DNA多态标记—小卫星和微卫星DNA[J]．国外医学-遗传学分册，1992，15(4)，172-173
[29] 李瑞生，陈振文，等．近交系大鼠微卫星DNA与生化标记分析的比较研究[J]．中国实验动物学杂志，2002，12(4)：225-228
[30] Brant J. Bassam, Gustavo Caetano-Anollés, et al. Fast and sensitive silver staining of DNA in polyacrylamide gels[J].Analytical biochemistry, 1991, 196:80-83
[31] 张艳，张树义．微卫星方法简介[J]，动物学杂志，1999，34(2)：42-45
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |