PCR扩增近交系大鼠微卫星位点DNA多态性的研究

本实验选取大鼠7条染色体上的微卫星位点合成了10对引物,利用聚合酶链反应（PCR）扩增技术对国内北京和哈尔滨等4家单位提供伯6个品系（SHR、SHRSP、LEW、RCS、WKY和F344）的8个近交系大鼠群体进行了DNA多态性分析的研究。结果表明：9个微卫星位点具有显多态性;不同品系个体之间具有多态性;同一群体不同个体之间除SHR（哈）的SMST位点和WKY（哈）的AGT位点出现一定的差异,其他均没有差异;不同地区同一品系的不同个体之间也存在一定的差异。该方法能有效地对近交系与杂交系、品系与品系、品系与亚系加以区分。因此,本实验为开展近交系大鼠遗传作图、基因定位和为实验动物的遗传背景监测提供可靠的信息,为大鼠遗传基因的研究提供了一个快速简例、特异准确的方法。     

近交系小鼠微卫星DNA多态性的研究

随机选择位于小鼠不同染色体上的微卫星引物42对,用PCR技术对C3H、C57、BALB/c、DBA、TA2、T739、B615、BACB/c－nu－nu和SCID等9种实验室常用近交系小鼠微卫星DNA多态性进行了研究。结果显示有信息的40对引物中,9种近交系小鼠在各基因座上均出现一条清晰条带,28个基因座表现为多态性。其中D3Mit22、D7Nds1、D11Mit12、D12Nds2、D15Mit17、D16Mit3、D16Mit4基因座表现为显著多态性。T739、B615和TA2的遗传背景相近,其相似系数分别为90％和85％;其次为TA2、SCID和B615,其相似系数分别为80％和82.5％。结果表明所检测的小鼠符合近交要求,筛选出的引物能典型地反映9个近交系小鼠的品系特异性和遗传背景,可用于常规检测小鼠品系来源和遗传背景等。Abstract：Forty-two microsatellites DNA loci on different chromosomes in nine kinds of inbred strain mice including C3H,C57,BALB/c,DBA,TA2,T739,B615,BALB/c-nu-nu and SCID were investigated by PCR analysis.It showed that all these mice tested display single allelic gene band with forty pairs of informative primers.Twenty-eight loci are polymorphisms,among which the polymorphisms of D3Mit22,D7Nds1,D11Mit12,D12Nds2,D15Mit17,D16Mit3,and D16Mit4 loci are significant.The genetic background of T739 was similarity with that of B615 and TA2 ,the similarity indices were 90％ and 85％ respectively;and that of TA2 was similarity with SCID and B615,the similarity indices were 80％ and 82.5％.These results suggest that these mice tested meet the request of inbred strain.Screened primers showing marked polymorphisms topically reflect the speciality of strains and genetic backgrounds,which could be used in determining the strains origin and genetic background of mice.     

近交系小鼠微卫星DNA多态性的研究

随机选择位于小鼠不同染色体上的微卫星引物42对,用PCR技术对C3H、C57、BALB/c、DBA、TA2、T739、B615、BACB/c－nu－nu和SCID等9种实验室常用近交系小鼠微卫星DNA多态性进行了研究。结果显示有信息的40对引物中,9种近交系小鼠在各基因座上均出现一条清晰条带,28个基因座表现为多态性。其中D3Mit22、D7Nds1、D11Mit12、D12Nds2、D15Mit17、D16Mit3、D16Mit4基因座表现为显著多态性。T739、B615和TA2的遗传背景相近,其相似系数分别为90％和85％;其次为TA2、SCID和B615,其相似系数分别为80％和82.5％。结果表明所检测的小鼠符合近交要求,筛选出的引物能典型地反映9个近交系小鼠的品系特异性和遗传背景,可用于常规检测小鼠品系来源和遗传背景等。Abstract：Forty-two microsatellites DNA loci on different chromosomes in nine kinds of inbred strain mice including C3H,C57,BALB/c,DBA,TA2,T739,B615,BALB/c-nu-nu and SCID were investigated by PCR analysis.It showed that all these mice tested display single allelic gene band with forty pairs of informative primers.Twenty-eight loci are polymorphisms,among which the polymorphisms of D3Mit22,D7Nds1,D11Mit12,D12Nds2,D15Mit17,D16Mit3,and D16Mit4 loci are significant.The genetic background of T739 was similarity with that of B615 and TA2 ,the similarity indices were 90％ and 85％ respectively;and that of TA2 was similarity with SCID and B615,the similarity indices were 80％ and 82.5％.These results suggest that these mice tested meet the request of inbred strain.Screened primers showing marked polymorphisms topically reflect the speciality of strains and genetic backgrounds,which could be used in determining the strains origin and genetic background of mice.     

微卫星DNA监控大鼠近交系的培育

采用微卫星DNA技术来监控大鼠仔代基因状况,选择性地进行交配繁殖,使基因快速纯合,缩短培育新的近交系动物周期。利用PCR扩增30个微卫星DNA位点对封闭群SD和Wistar大鼠交配繁殖的仔代鼠进行微卫星DNA多态性分析,仔代中与母代SD大鼠相似系数高的与中的进行定向交配繁殖。F2代大鼠均为杂合多态的位点,没有纯合位点;到F9代时基因纯合位点达27个,纯合基因位点率为90%。每代相似系数具有不断上升的趋势,上升率为6-20%。采用皮肤移植方法验证了F9代大鼠间无排斥。建立了一种新的快速培育近交系动物的方法。     

3个山羊群体中4个微卫星DNA多态性及其与杂种优势的关系

利用4个微卫星标记（OarFCB11,OarAE101,McM218,McM38）对波尔山羊、太行山羊和河北奶山羊的等位基因频率、群体多态信息含量、有效等位基因数、杂合度和遗传距离进行了遗传检测,并测定了波尔山羊与河北奶山羊及太行山羊的杂交效果。结果表明：4个微卫星标记在波尔山羊、太行山羊和河北奶山羊3个品种中存在多态性,可以用于山羊遗传多样性的评估;从不同品种来看,太行山羊的遗传变异程度最大,而波尔山羊的遗传变异程度相对较小;波尔山羊与河北奶山羊的遗传距离大于与太行山羊,波尔山羊与河北奶山羊的杂种优势高于与太行山羊,与实际杂种优势测定结果相符。Abstract: Gene frequency, polymorphism information contents, number of effective alleles, heterozygosity and genetic distances were studied in Boer goat, Taihang goat and Hebei dairy goat using four microsatellite markers（OarFCB11,OarAE101,McM218,McM38）. The crossing effects on Hebei dairy goat and Taihang goat with Boer goat were tested. The results indicated that there are genetic polymorphisms at four microsatellite markers in three goat breeds. Four microsatellite markers can be used for genetic diversity evaluation in goat breeds. The genetic variability of Taihang goat is the highest, and Boer goat is the lowest in three goat breeds. Genetic distances between Boer goat and Hebei dairy goat is bigger than that between Boer goat and Taihang goat. The heterosis between Boer goat and Hebei dairy goat is higher than that between Boer goat and Taihang goat. It accords with testing results on actual heterosis.     

猪微卫星标记多重PCR扩增组合

采用多重PCR的方法,以快速扩增微卫星标记和节约试剂为目标,对其反应条件进行优化后,获得了46个扩增效果理想的微卫星标记多重PCR组合,其中30个为二重PCR,16个为三重PCR。实验结果表明,这些多重PCP反应的引物浓度为0.06~0.3μmol/L, Mg2+的浓度变化范围为1.5~3.0mmol/L,采用的PCR缓冲液的倍数为1.0、1.2、1.4或1.6,每个PCR反应聚合酶的用量在0.2和0.4U之间,退火温度及反应循环数分别为52~60℃ 和32~50℃。所有多重PCR进一步合并为17个可在ABI 337测序仪上进行电泳的组合。Abstract:In order to rapidly amplify pig microsatellite markers and save materials,multiplex PCR was used and its reaction condition was optimized.Forty-six combinations of multiplex PCR with good effects were obtained.Thirty of them are duplex-PCRs,sixteen are triplex-PCRs.The results of multiplexes showed that the concentration of primers varied among 0.06~0.3μmol/L,the Mg2+ concentration among 1.5~3.0 mmol/L;0.2~0.4 U of Taq polymerase and 1.0-,1.2-,1.4-,1.6-fold buffer were used,the annealing temperature and the cycle number varied among 52~60℃ and 32~50℃,respectively.All multiplexes were further combined into 17 sets for the electrophoresis on ABI 377 sequencer.     

树鼩微卫星DNA的多态性研究

目的探索并建立一种检测树鼩群体遗传多样性的方法。方法利用聚合酶链反应（PCR）扩增技术对30只树鼩个体的11个微卫星位点进行了遗传检测。结果所选的11个微卫星DNA位点中,有9个具有高度多态性,2个微卫星DNA位点多态性较差。结论所研究的树鼩微卫星位点中,有9个符合遗传标记特点,可用于检测树鼩群体的遗传多样性。     

海参微卫星DNA的多态性

微卫星DNA(microsatellite DNA)广泛存在于真核生物的基因组中。由于其具有突变频率快、多态性丰富、呈共显性遗传、通用性等特点,已成为近年来被广泛应用的分子遗传标记。本研究对10个海参微卫星DNA进行了克隆与测序。结果表明:90%的微卫星DNA序列存在长度多态性,这为进一步研究海参的分子标记辅助育种奠定了基础。     

应用40对微卫星引物对6种近交系小鼠遗传背景进行监测的研究

采用40对引物的微卫星DNA PCR遗传质量符合要求的BALB／C－nu0nu-,DBA/2,SCID,T739,TA2,615等6种近交系小鼠进行遗传监测,结果26对引物有稳定的扩增结果,5对引物表现为单态性,21对引物表现出多态性,其中D2Nds3,D3Mitl5,D3Mitl7,D3Mit18,D16Mit7等6对引物表现出显著的多态性,反映了各品系小鼠独特的遗传背景,可应用于区别小鼠品系,监测系间的遗传污染,为有关小鼠品系积累了遗传背景资料,有助于将实验动物的遗传监测从表墼这度到DNA水平。     

用DNA指纹图技术分析Wistar大鼠的遗传距离

采用JL-02多位点探针和Southern杂交对Wistar大鼠基因组进行了DNA指纹分析,并对国内最具有典型的6个地区9个Wistar大鼠群体内和群体间进行了DNA指纹分析比较。结果表明,DNA指纹图较好地反映了封闭群动物的遗传本质,具有良好的多态性。同一群体不同个体间Wistar大鼠遗传距离主要为0.2~0.6,将不同群体的Wistar大鼠DNA指纹图带进行分析表明,所有群体间的DNA指纹图的遗传距离在0.2~0.7。Abstract:DNA fingerprinting of Wistar rat were studied with JL-02 Mulilocus probe and Southern hybridization,and comparing with different individuals of nine groups Wistar rat from six national classical area and different groups.It was indicated that DNA fingerprinting could reflect genetic material of outbred strain rat,and were more polymorphic.The genetic distances of Wistar rat were distributed for 0.2~0.6 among different individuals within same groups,and the genetic distances of Wistar rat were distributed for 0.2~0.7 among different groups．