人类疾病模型小鼠的标准化维持方法研究

目的研究人类疾病模型小鼠的可遗传生物学特性,建立品系的标准化维持方法。方法选用FVB TgN、MRL Mpj和SAMP1Ka三个疾病模型小鼠品系作为代表性实验对象,通过测定生长曲线、RAPD同工、酶电泳、行为学试验等方法,找出三个品系相对于各自对照品系的不同特征,并建立了标准化检测指标作为维持方法的依据。结果MRL Mpj的生长曲线相对于对照品系有明显的统计学差异;FVB TgN、MRL Mpj、SAMP1Ka等三个品系的RAPD图谱在阳性引物及扩增出的条带方面均不一致;同工酶电泳的结果表明不同品系的个体之间表型不完全相同;行为学试验更从多方面直观地显示了各品系的不同特征。结论人类疾病模型小鼠除了用常规检测方法外,还应建立各品系在生长曲线、RAPD、同工酶电泳、行为学试验等方面的特殊检测方法,及时检测模型小鼠品系的独特性状并作为保种依据,以避免遗传漂变的发生。     

近交系小鼠遗传质量监测新方法的建立——染色体Q—H分带技术及其应用

本研究利用最新的Quinacrine Mustard and 33258 Hoechst(Q—H)复合荧光染色技术对10个品系的近交系小鼠的核型进行分析。在同一细胞内,按各号染色体着丝粒带大小排列、分组,建立该10个品系近交系小鼠特有的染色体着丝粒带核型,作为各品系小鼠遗传质量监测的染色体标记指标。本研究还对615小鼠品系的生化标汜检测与染色体标记检测的结果进行比较,同时比较了不同来源615小鼠的染色体标记,从而进一步阐明了该方法作为实验动物遗传监测方法之一与其他方法间的互补性及其自身特点。     

巴马小型猪与贵州小型香猪遗传多样性的RAPD分析

应用经过筛选的31条引物对巴马小型猪和贵州小型香猪的基因组DNA进行RAPD扩增,分析二品系实验用小型猪的遗传多样性.两品系实验用小型猪品系内及品系间多态性位点的百分数分别为30.9%和25.7%,品系间及品系内的平均遗传距离分别为0.120、0.072和0.067.结果表明,两品系实验用小型猪品系间及品系内遗传多样性贫乏,遗传变异较小.     

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。     

NJS小鼠与其亲本小鼠遗传多样性的RAPD分析

目的分析NJS及其亲本小鼠的遗传结构.方法筛选出5条随机引物对NJS及其亲本小鼠的遗传结构进行RAPD分析,并对NJS小鼠个体之间的基因组DNA进行相似性分析.结果两种小鼠均有相同的扩增片段且产生了各自的特异性条带;NJS小鼠不同个体间拥有的RAPD条带也具有差异,但拥有相同条带的个体小鼠比率较高.该群体小鼠的相似系数(F)为0.927(0.880～0.980).结论 RAPD分析获得的多态性可用于NJS与其亲本小鼠之间的遗传分析;而且NJS小鼠个体间具有较好的遗传一致性和遗传稳定性,其群体分化程度处在一个较低的水平.     

利用多重荧光STR技术分析上海地区7品系常用近交系小鼠核心群的遗传特性

目的比较上海地区7品系常用近交系小鼠核心群的遗传特性。方法将筛选到的48对多态性丰富的微卫星引物组合优化,形成11组多重荧光PCR引物混合体系,对来自上海地区两大实验小鼠供应商的7品系近交系小鼠核心群的DNA样进行分型检测。利用遗传分析软件进行数据分析。结果来自两大供应商的7品系近交系小鼠在48个微卫星位点上都为纯合子。同一种群内小鼠的STR位点结果均一致;不同种群小鼠无论品系是否相同,相互间均存在STR位点差异。但相同品系不同种群近交系小鼠间的遗传距离与不同品系小鼠种群间的遗传距离相比均较近。在UPGMA聚类树中,相同品系的不同种群均首先两两聚成一类。C57BL/6小鼠与其他6品系小鼠的亲缘关系均较远。结论上海地区不同供应商的7品系近交系小鼠核心群间均存在STR位点差异。     

科技动态

动物毛发异常对行为学研究的影响动物游泳试验被广泛的应用于研究啮齿类动物行为学研究。暴露于水中的动物毛发和皮肤是影响实验结果的重要因素。经过遗传修饰或药物处理过的实验动物,经常会出现毛发的异常。在对这些动物进行游泳试验时,就可能会影响到试验结果。芬兰的Kalueff AV等人为了探求毛发异常在动物强迫游泳试验中的作用,他们将129S1品系的实验小鼠脱毛并和未脱毛的小鼠对比它们在游泳中表现的不同。实验结果显示,脱毛小鼠在5分钟的强迫游泳实验中没有表现出与未脱毛小鼠有何不同之处,认为该品系的小鼠毛发改变对于游泳结果不是…     

BALB/c小鼠遗传质量检测的新方法

目的比较随即扩增多态性方法（RAPD）、微卫星方法（STR）与生化标记方法对近交系小鼠遗传质量检测的差异,为近交系动物遗传质量控制提供一种分子生物学方法。方法提取近交系小鼠BALB/c基因组DNA,用6条RAPD引物和20对STR引物对其进行PCR扩增,用生化标记法检测13个位点。结果在6条RAPD引物中,引物2（p2）、引物3（p3）、引物5（p5）和引物6（p6）这四条引物扩增的条带出现差异,表现为不同的RAPD图谱;在20对STR引物中,引物2、4、10和11,这四对引物扩增的条带出现差异,表现为不同的STR图谱;13个生化标记位点中,过氧化氢酶-2（Ce-2）等6个生化位点发现杂合基因。结论RAPD和STR可用于验证生化标记方法的实验结果,并用于保证近交系动物的遗传质量。     

双色荧光杂交芯片在近交系小鼠遗传监测中的应用

应用一种新的高通量SNP检测方法-双色荧光杂交芯片技术进行近交系小鼠遗传监测。应用双色荧光杂交芯片技术对4个品系近交系小鼠的多个基因组DNA样本进行SNP分型,整合6个SNP位点的芯片杂交信息,对样本所属品系进行判断。研究结果表明SNP检测方法-双色荧光杂交芯片技术能够对选定的6个SNP位点进行高准确率分型;双色荧光杂交芯片技术是一种高通量SNP检测的良好工具,适合于对少量近交系品系来源的大样本量小鼠进行遗传污染监测和品系鉴定,并具有扩大应用的潜力。     

双色荧光杂交芯片在近交系小鼠遗传监测中的应用

应用一种新的高通量SNP检测方法-双色荧光杂交芯片技术进行近交系小鼠遗传监测。应用双色荧光杂交芯片技术对4个品系近交系小鼠的多个基因组DNA 样本进行SNP分型,整合6个SNP位点的芯片杂交信息,对样本所属品系进行判断。研究结果表明SNP检测方法-双色荧光杂交芯片技术能够对选定的6个SNP位点进行高准确率分型;双色荧光杂交芯片技术是一种高通量SNP检测的良好工具,适合于对少量近交系品系来源的大样本量小鼠进行遗传污染监测和品系鉴定,并具有扩大应用的潜力。     

一种新的近交系实验动物遗传监测方法及小鼠性别相关RAPD标记的发现

我们用21个10 bp的随机短引物对来自昆明、成都、上海、北京四个地方的BALB/c小鼠以及C57BL小鼠、昆明种小白鼠进行了随机扩增多态DNA(RAPD)分析,发现13个引物的扩增产物在BALB/c小鼠和C 57 BL小鼠中有差异,8个引物的扩增产物在BALB/c小鼠和昆明种小白鼠之间不同.在四个地方的BALB/c小鼠中,成都、上海、北京的BALB/c小鼠其遗传背景均一,而来自昆明的BALB/c小鼠中,有2只的4个引物的扩增产物不同于其它的BALB/c小鼠,表明这两只BALB/c小鼠可能曾发生过某种程度的遗传改变或污染。实验结果显示RAPD方法是一种有效的近交系实验动物遗传监测手段。实验中一个有趣的结果是,在OPG 2、OPE 4、OPE 9的扩增产物中,发现了严格的性别依赖的PAPD标记。OPE 9扩增产物中,凡雄性个体都有一条0.88 kb的标记.OPG 2、OPE 4则在所有的雄性个体中多扩增出一条约1.2 kb的带。通过交叉PCR扩增和斑点杂交证明OPG 2、OPE 4 得到的雄性特异性RAPD标记虽分子大小一致,但不具同源性。这些性别相关RAPD标记的染色体定位和性质分析正在进一步进行中.     

辽宁省6种常用近交系小鼠10个微卫星位点的遗传质量检测报告

目的利用微卫星技术对辽宁省6种近交系小鼠进行遗传质量分析。方法根据Mouse Genome Database和相关文献选取10个多态信息丰富的位点和引物,进行PCR扩增和PAGE电泳,对小鼠的遗传多态性进行研究。结果不同品系小鼠同一位点的扩增结果表现出多态性,同一品系同一位点表现单态性,所有小鼠的10个位点都处于纯合状态;遗传距离分析表明,C57BL/10与C57BL/6J小鼠之间的遗传距离最近,为0.1021,遗传距离最远的是BALB/c与C57BL/10、C57BL/6J,分别为0.1635和0.1614。结论运用所筛选的10个微卫星位点可以对近交系小鼠进行遗传质量检测,说明该方法具备可行性。     

Characteristics of mouse lymphoid cells proliferating in vitro by recombinant human interleukin 2 (TGP-3): comparison between normal and nude mice

Various lymphoid cells obtained from BALB/c and BALB/c nu/nu mice were cultured in vitro with recombinant human interleukin 2 (rIL 2), and the characteristics of responder cells to rIL 2 were analyzed. Spleen cells, lymph node cells, and thymocytes except for bone marrow cells obtained from BALB/c mice remarkably proliferated in response to rIL 2. On the other hand, among lymphoid cells obtained from BALB/c nu/nu mice, only lymph node cells showed significant proliferation by rIL 2. Flow cytometric analysis revealed that mainly two types of lymphoid cells were proliferating in response to rIL 2 in BALB/c mice, i.e., Thy 1+, Lyt 1-, Lyt 2- and Thy 1+, Lyt 1-, Lyt 2+ cells. On the other hand, most of the proliferating cells were Thy 1+, Lyt 1-, Lyt 2- cells in BALB/c nu/nu mice. Treatment with various antibodies plus complement revealed that the majority of IL 2-responsive cells in BALB/c mice were Thy 1+, Lyt 1+, and Lyt 2+, although a minor part of them were Thy 1-, Lyt 1-, and Lyt 2-. On the other hand, a predominant type of the IL 2-responsive cells in BALB/c nu/nu mice were Thy 1-, Lyt 1-, and Lyt 2-, though some were Thy 1+. Nonspecific killer activity against tumor cells increased to variable extents in all of the lymphoid cells of both strains after culture with rIL 2. Our results indicate that mouse responder cells to rIL 2 have the following characteristics. First, the responder cells exist abundantly among spleen, lymph nodes, and thymus in normal mice, though their cell lineages are heterogeneous; one is of T cell lineage and the other of natural killer (NK) cell lineage. Second, nude mice are defective in the responder cells of T cell lineage but not of NK cell lineage. Moreover, the responder cells in nude mice predominantly accumulate in the lymph nodes but not other lymphoid organs.     

Host defense in cryptococcosis. II. Cryptococcosis in the nude mouse.

In the homozygous state, mice carrying the “nude” (nu) gene are hairless (nude), lack a thymus and have profound deficiency of cell-mediated immunity. Cryptococcosis was studied in BALB/c and Swiss mice, each strain carrying the nu gene. The purpose was to determine the interactions of the nu gene and mouse strain in terms of susceptibility to Cryptococcosis. Mice of both strains could be sensitized to produce delayed-type hypersensitivity reactions to cryptococcal extract in the heterozygous nu/X state, but not in the nu/nu state. Nu/X Swiss mice were more resistant than nu/X BALB/c mice to infection with a highly virulent strain (B) of Cryptococcus neoformans. However, nu/nu BALB/c and nu/nu Swiss mice were both highly susceptible to the same microorganism. Challenge with another cryptococcal strain (A) of much lower virulence for nu/X mice killed 100% of BALB/c and Swiss nu/nu mice. These studies indicate that thymus-dependent immune functions are critical determinants of host resistance to murine Cryptococcosis.     

人工雌核发育鲢的遗传多样性及异源遗传物质整入的RAPD分析

运用RAPD技术对连续二代人工雌核发育鲢的遗传多样性及异源遗传物质的整入进行了分析 ,结果表明 :一代雌核发育鲢 ,个体间遗传相似度为 0 94 5— 0 995 6 ,多样性指数为 0 175 ;二代雌核发育鲢 ,个体间遗传相似度为0 96 15— 1 0 0 ,平均为 0 985 2 ,多样性指数为 0 0 6 2。研究揭示经过连续二代人工雌核发育后 ,其遗传多样性明显减少 ,种质进一步纯化。通过对雌核发育鲢二代、亲本鲢和雄鲤的RAPD扩增比较 ,发现雌核发育鲢含有少数与父本相同的特异DNA扩增带 ,而亲本鲢没有 ,在基因水平上表明雌核发育鲢整入了雄鲤的遗传物质     

Cells with natural killer activity in the cerebrospinal fluid of normal mice and athymic nude mice with acute Sindbis virus encephalitis

Sindbis virus causes an acute, nonfatal inflammatory encephalitis in weanling BALB/c mice. Mononuclear inflammatory cells are present in the cerebrospinal fluid (CSF) as well as in the parenchyma of the brain. Both aspects of this inflammatory response were eliminated by treatment with cyclophosphamide. Athymic nude (nu/nu) mice developed no inflammation in the brain, but did develop a CSF pleocytosis that peaked on day 2 after infection. The time course of the appearance of cells in the CSF was earlier in nu/nu mice than their heterozygote (nu/+) littermates. The pleocytosis in nu/nu mice reached a peak on day 2, whereas in nu/+ mice the peak was on day 4, as it is in normal BALB/c mice. To determine whether some of the CSF cells in nu/nu mice may be natural killer (NK) cells, NK activity was measured in a 4-hr assay by using a YAC-1 target cell. NK cell activity in the spleen and peripheral blood was induced by infection with Sindbis virus in nu/nu mice with a similar time course to that of nu/+ mice (peak 1 day after infection). CSF from nu/nu mice had NK activity present 2 days after infection that was greater than that present in either the peripheral blood or spleen. BALB/c and nu/+ mice had insufficient cells present for assay at day 2, but BALB/c mice had NK activity present in the CSF 3 and 5 days after infection that exceeded that in the peripheral blood or spleen. Brain interferon was detectable on day 1 in nu/nu mice, but not until day 2 in nu/+ mice even though the amounts of brain virus were the same in the two groups at all time points. It is concluded that cells with NK activity contribute to the CSF pleocytosis induced by acute Sindbis virus encephalitis.     

河南地方野生小鼠与近交系小鼠的扩增片段长度多态性分析

目的 从分子水平上阐明河南省兰考地区捕获的野生小鼠(LK)与4个小鼠品系(B6,BALB/c,DDK,PWK)之间的遗传差异及遗传关系,从而进一步确定该种野生小鼠的种属及培育野生来源近交系小鼠品系.方法 利用25对引物,对野生小鼠及4个小鼠品系进行扩增片段长度多态性(AFLP)分析.结果 检测到2035条扩增条带,多态...     

新疆吐鲁番荒漠土壤绿藻的多样性分析

应用RAPD技术对吐鲁番地区火焰山及艾丁湖区域分离的15株土壤绿藻(chlorophyta)品系的遗传多样性及其亲缘关系进行探讨。结果表明:从20个随机引物中,筛选出多态性和重复性较好且谱带清晰的引物8个,这8个引物扩增出的DNA片段大多在300～2 000 bp之间,所形成的多态性位点数差距较大,显示该区域土壤绿藻具有较丰富的遗传多样性;15株土壤绿藻扩增共得到74条谱带,71条多态性带,其多态性比率为95.95%;聚类分析显示15株土壤绿藻明显地聚为2大类,与其来源相对应,即隶属于同一亚组或相近亚组的不同种基本归为一类,其种间关系与传统的形态学分类结果相吻合。     

Increased membrane immunoglobulin capping of B cells from C57Bl/6 lpr/lpr and C57Bl/6 nu/nu mice

When the capping of membrane immunoglobulin on spleen B cells from normal C57Bl/6 mice (B6) is taken as reference, a faster capping rate is found for cells of age-matched B6 mice which are congenic at the lymphoproliferation (lpr) or nude (nu) loci. Though both congenic strains can be characterized by an abnormal T-lineage cell content, the nature of the abnormality itself is very different since B6 nudes lack thymus-processed/influenced lymphocytes whereas B6 mice with the lpr phenotype suffer from an invasion of all lymphoid organs with cells of a particular T-cell subset. Moreover, the more "normal" capping rate of B cells from the double congenic B6 mice (nu/nu, lpr/lpr) is intriguing. Since other mice homozygous at the lpr locus (MRL-1) or at the nu locus (BALB/c nude) also cap faster than their congenic controls (MRL-n and BALB/c, respectively), the observed effects do not appear to depend on a peculiarity of the B6 genetic background. If the faster capping of B cells of nu congenic and of lpr congenic mice had a common origin, it might be that T cells would control in some way the mobility of B-cell membrane immunoglobulins: both congenic mice have in their spleen a very low proportion of mature T cells together with a very high proportion of prethymic/thymic immature T-cell types, either of which might affect B-cell behavioral responses to membrane immunoglobulin clustering.