血清淀粉样P成分转基因小鼠模型的建立

目的:建立可表达血清淀粉样P成分(SAP)转基因小鼠模型,以研究SAP基因的功能。方法:采用基因重组技术将SAP基因插入到pCMV-Tag5A真核表达载体上。经BciVI限制性内切酶酶切后,琼脂糖凝胶电泳回收2.7 kb目的片断,受精卵原核显微注射法将其注入雄原核,制备转基因小鼠。采用PCR法在基因水平筛选SAP转基因小鼠阳性小鼠;免疫沉淀法结合免疫印迹法在蛋白水平检测SAP基因在PCR阳性转基因小鼠中的表达情况。结果:成功构建了pCMV-Tag5A/mSAP真核表达载体。经显微注射法将2.7 kb线性目的基因片断注射入受精卵雄原核,移植入代孕母鼠,所生127只小鼠,其中PCR鉴定获得10只转基因阳性小鼠。免疫沉淀法和免疫印记法随机检测83#♂founder、7#♀founder与C57 BL/6J小鼠回交F1代PCR阳性转基因小鼠血清中外源SAP基因蛋白水平的表达,发现均有表达。结论:该研究以C57BL/6J小鼠为研究对象,成功地产生能稳定遗传SAP基因的过表达转基因小鼠模型,它将有利于研究SAP基因在炎症、淀粉样沉淀、自身免疫系统中的作用。     

SPRN转基因小鼠感染Scraipe鼠适应性病毒后动物行为与脑组织形态比较

通过建立Scraipe鼠适应性病毒感染Tg(sprn-B)/C57BL/6转基因小鼠与C57BL/6野生型小鼠,观察鼠行为学及脑组织形态学的病理变化,探讨Shadoo蛋白在感染过程中对小鼠学习记忆能力及中枢神经系统的影响。结果显示在Morris水迷宫定位航行中,4~7 d转基因感染组与野生型感染组的逃避潜伏期差异极显著(P<0.01);空间探索试验中,转基因感染组与野生型感染组的第1次穿越平台时间及穿越平台次数差异极显著(P<0.01)。HE染色观察鼠脑切片发现,转基因感染组与野生型感染组相比,海马、皮质、小脑空泡数量略少,海马区锥体层细胞排列比较紧密,细胞轮廓比较完整,小脑蒲肯野氏细胞清晰明显。     

Prevention of allogeneic bone marrow graft rejection by H-2 transgene in donor mice

Rejection of bone marrow grafts in irradiated mice is mediated by natural killer (NK) cells and is controlled by genes linked to the major histocompatibility complex (MHC). It has, however, not been possible to identify the genes or their products. An MHC class I (Dd) transgene introduced in C57BL donors prevented the rejection of their bone marrow by NK cells in irradiated allogeneic and F1 hybrid mice expressing the Dd gene. Conversely, H-2Dd transgenic C57BL recipients acquired the ability to reject bone marrow from C57BL donors but not from H-2Dd transgenic C57BL donors. These results provide formal evidence that NK cells are part of a system capable of rejecting cells because they lack normal genes of the host type, in contrast to T cells, which recognize cells that contain abnormal or novel sequences of non-host type.     

C57BL/6和A/J两品系小鼠脾脏消减cDNA文库的构建及差异基因分析

【目的】检测健康C57BL/6和A/J小鼠常规免疫指标,构建脾脏消减cDNA文库并筛选差异表达基因,以探讨两品系小鼠对猪链球菌抗病性差异的分子机制。【方法】利用ELISA法检测血清常规免疫指标,抑制性消减杂交（suppression subtraction hybridization,SSH）技术构建8周龄小鼠脾脏差异表达基因的cDNA文库。【结果】试验结果表明,A/J品系小鼠血清中IgA水平明显高于C57BL/6品系小鼠（P＜0.05）,而血清IgG和IFN水平在两品系小鼠间均无显著差异。对筛选的149个阳性克隆进行测序,去除冗余的cDNA序列载体并聚类拼接后获得56条差异表达序列标签（ESTs）。利用Genebank的BLAST分析核酸和蛋白质同源性比较,26个不同的基因或ESTs具有高度的同源性,2条ESTs未找到同源序列。【结论】筛选到很多EST与信号转导、细胞凋亡及免疫等重要功能基因高度同源,为研究猪链球菌的致病机理和防治提供了实验依据。     

Male-specific antigen: modification of potency by the H-2 locus in mice

Skin grafts from C57BL/10 (B10) male mice survive significantly longer than do B10.BR male skin grafts on (B10 x X B10.BR)F(1) females. This indicates that the H-2 locus influences the potency of the male-specific antigen in mice.     

Repression of colony formation reversed by antiserum to mouse thymocytes

Marrow cells derived from C57BL/6 mice form many fewer splenic colonies in irradiated C57BL/6 x C3H F1 hybrid recipients than in irradiated C57BL/6 recipients (repression of colony formation). This effect is reversed by treatment of the hybrid recipients with active antiserum to mouse thymocytes. The repression phenomenon cannot readily be explained in immunological terms; hence the effect of the antilymphocyte serum on this phenomenon may not result from immunosuppression in the usual sense.     

鲤疱疹病毒Ⅱ型(CyHV-2) ORF25蛋白的原核表达及单克隆抗体的制备

利用PCR方法扩增鲤疱疹病毒Ⅱ型(CyHV-2)ORF25基因的部分基因序列,并将扩增产物连接到原核表达载体pGEX-KG,成功构建了CyHV-2-ORF25-KG重组质粒。IPTG将其诱导后,SDS-PAGE分析表明:目的蛋白得到表达,分子质量大小为42ku,且主要以包涵体形式存在;用纯化的融合蛋白免疫BALB/c小鼠,多次免疫后通过细胞融合,筛选出1株单抗5C6;经间接免疫荧光试验和Western blot试验验证,这株单抗可以识别CyHV-2ORF25蛋白。     

Specific expression of hepatitis B surface antigen (HBsAg) in transgenic mice

Two transgenic mice were obtained that contain in their chromosomes the complete hepatitis B virus (HBV) genome except for the core gene. These mice secrete particles of HBV surface antigen (HBsAg) in the serum. In one mouse, HBV DNA sequences that had integrated at two different sites were shown to segregate independently in the first filial generation (F1) and only one of the sequences allowed expression of the surface antigen. Among these animals the males produced five to ten times more HBsAg than the females. A 2.1-kilobase messenger RNA species comigrating with the major surface gene messenger RNA is expressed specifically in the liver in the two original mice. The results suggest that the HBV sequences introduced into the mice are able to confer a tissue-specific expression to the S gene. In addition, the HBV transgenic mice represent a new model for the chronic carrier state of hepatitis B virus infection.     

Depletion of conventional mature B cells and compromised specific antibody response in bovine immunoglobulin μ heavy-chain transgenic mice

In this study, we introduced the bovine immunoglobulin μ heavy-chain gene (the orphaned gene on BTA11) into mouse germline cells. Bovine IgM was highly expressed in selected transgenic lines, and it largely inhibited rearrangements of the endogenous immunoglobulin heavy chain (IgH) genes in these lines. The forced expression of bovine IgM resulted in reduced numbers of pro- and pre-B cells but increased the number of immature B cells in the transgenic mice. Bovine IgM-expressing B cells can migrate from the bone marrow to the spleen, but most of the cells are arrested at the T1 transitional B cell stage, leading to a significantly lower number of T2 transitional and mature B cells in the spleen. Although the serum concentrations of endogenous IgM and IgG in the transgenic mice were significantly decreased, the IgA levels were slightly increased compared to the WT mice. The bovine IgM level in the serum was only one-tenth to one-fifth of that of endogenous mouse IgM, suggesting that most of the serum immunoglobulin were contributed by endogenous IgH gene-expressing B cells. These transgenic mice also exhibited a lower frequency of unique complementarity determining region 3 (CDR3) sequences in their VH repertoire and Vκ repertoire but exhibited an increased frequency of unique CDR3 in their Vλ repertoire. Compared to the WT mice, the transgenic mice had a significantly higher percentage of mouse IgM-expressing B cells that expressed λ chains. Finally, we showed that the transgenic mice were deficient in a specific antibody response to antigen stimulation.     

周龄·初始注射时间对小鼠超数排卵的影响

[目的]探讨不同周龄、不同初始注射时间对小鼠超数排卵效果的影响。[方法]用10 IU PMSG+10 IU HCG间隔48 h处理C57BL/6J小鼠,收集并统计合子数。[结果]4周龄超排效果优于于8周龄,可达26.86枚∕只;4周龄C57BL/6J小鼠PMSG和HCG注射时间为12:00—13:00,超排后收集合子数可达30枚∕只以上,而且体外培养的发育状况良好。[结论]4周龄C57BL/6J小鼠适宜超数排卵;PMSG和HCG注射时间为12:00—13:00,超排效果相对理想。     

H-2-linked resistance to mastocytoma in male mice: immune response to a histocompatibility antigen on the X chromosome

Male hybrids from a cross between female mice of strain C57BL/6Kh and males of strain DBA/2J lived longer after injection of P815 mastocytoma cells of DBA/2 origin than did their female siblings. Responses to the histocompatibility antigen on the X chromosome of the DBA/2 strain may be involved in resistance to the tumor. When the female parent was replaced with a C57BL/6Kh carrying one of several mutations in the H-2 region, this sex effect disappeared in some of the hybrid combinations. Thus, the H-2 complex appears to be involved in the regulation of the immune response to the X-linked histocompatibility antigen in this tumor model.     

Infection of the basal ganglia by a murine coronavirus

The coronavirus, mouse hepatitis virus strain A59 (MHV-A59), causes mild encephalitis and chronic demyelination. Immunohistochemical techniques showed that MHV-A59-infected C57BL/6 mice contained dense deposits of viral antigen in the subthalamic nucleus and substantia nigra, with fewer signs of infection in other regions of the brain. The animals showed extra- and intracellular vacuolation, neuronal loss, and gliosis in the subthalamic-nigral region. Such localization is unprecedented among known viral encephalitides of humans and other species. This infection by a member of a viral class capable of causing both encephalitis and persistent infection in several species may be related to postencephalitic parkinsonism.     

年龄和性别对C57BL/6小鼠种属型行为的影响

探讨C57BL/6小鼠年龄和性别对其种属型行为的影响.结果表明,老年小鼠的挖掘行为无显著性损害,但老年小鼠的作窝能力显著下降.老年和成年雄鼠的挖掘行为均较同龄雌鼠有显著性下降,老年雄鼠的作窝能力显著差于同龄雌鼠.储藏实验显现出无论是老年还是成年C57BL/6小鼠均无明显的储藏行为.这些结果提示作窝能力是反映老年C57BL/6小鼠衰老效应及性别效应的良好指标,挖掘行为可以判别各年龄C57BL/6小鼠的性别效应,储藏行为无实际意义.     

Deficiency of a kidney metalloproteinase activity in inbred mouse strains

Kidneys from BALB/c mice contain a potent metalloendoproteinase, termed meprin, that is active against large proteins as well as small peptides. The enzyme is present in mouse strains C57BR/cdJ, C57BL/6J, BALB/cJ, A/J, DBA/IJ, CD/l, Swiss, and ICR. Three related inbred strains, CBA/J, CBA/CaJ, and C3H/He, are markedly deficient in this enzymatic activity. This is the first report of a heritable deficiency of an intracellular proteinase in mammalian tissues. Meprin deficiency appears to have arisen as an early event in the development of the C stock. Furthermore, meprin is present in the progeny of a cross between a meprin-sufficient female (C57BL/6) and a meprin-deficient male (C3H/HeN), an indication that the trait for the deficiency is recessive.     

中年C57BL/6和ICR小鼠感觉运动能力的改变

探讨中年C57BL/6和ICR小鼠感觉运动能力的改变及实验装置对结果的影响.结果表明在10 mm和14 mm直径钢质平衡杆上,中年C57BL/6小鼠的平衡时间缩短,且雌鼠和雄鼠在10 mm直径钢杆上均可显示年龄效应.中年C57BL/6小鼠在紧绳实验中的悬挂时间和转换分也减少,但仅转换分在雄鼠显示年龄效应;中年ICR鼠在6 mm和14 mm直径钢杆上的平衡时间延长,且雌鼠和雄鼠在2种钢杆上均可显示年龄效应,但紧绳任务的悬挂时间和转换分均无年龄和性别差异.这些结果提示中年小鼠有感觉运动能力改变,但改变模式因任务和品系而异,平衡木的大小和形状影响年龄效应检测的有效性.     

Rapid immunological induction of murine lymphomas: evidence for a viral etiology

A graft-versus-host reaction induced in (SJL/J x C57BL/1)F(1) hybrid mice by injection of SJL/J spleen cells resulted in 100 percent incidence of tumors at 40 days. Transplantation studies revealed that the tumors were antigenically C57BL/1. Since both SJL/J and C57BL/1 mice carry tumorigenic virus, the evidence suggests a viral etiology.     

5种不同商品化的饲料对C57BL/6J和KM小鼠生长和繁殖的影响

研究市场上常见的5种饲料对C57BL/6J小鼠及KM小鼠生长和繁殖的影响,为实验小鼠生产中合理使用饲料提供科学依据.实验选取3周龄C57BL/6J小鼠和KM小鼠各250只,每种鼠随机分为5组,每组50只,雌雄各半,饲喂A,B,C,D,E共5种不同厂家的实验动物维持饲料,测定小鼠4～8周龄的体质量和脏器系数.另选取C57BL/6J小鼠及KM小鼠性成熟小鼠各150只,雌雄各半,每种鼠随机分为5组,每组15对,饲喂A,B,C,D,E共5种不同厂家的实验动物繁殖饲料,测定母鼠的产仔数、连胎率.结果表明:①5种维持饲料均能满足小鼠的生长需求, C饲料和E饲料更符合小鼠生长的营养需求.②5种饲料饲喂的小鼠器官指数正常.③5种繁殖饲料饲喂的母鼠第1,2胎的产仔数差异不具有统计学意义(p0.05),说明产仔数更多取决于遗传因素,本次调查的几种饲料对其影响较小.合理选择实验小鼠所需饲料,利于实验数据的稳定.     

基础研究BXSB小鼠肾组织NF-κB的表达及意义

目的:了解NF-κB在BXSB狼疮性肾炎小鼠肾组织中的表达及意义。方法:取BXSB狼疮性肾炎小鼠(即LN组)与C57BL/6小鼠(即对照组)各10只,应用免疫组织化学SP法检测肾组织中NF-κB的表达,并与病理学指标行等级相关分析。结果:LN组小鼠肾组织中NF-κB表达较对照组显著增高(P<0.05);NF-κB在肾小球和肾小管均有表达,但以肾小管表达更为显著;肾小球NF-κB阳性细胞数、NF-κB阳性肾小管数与肾组织活动指数呈高度正相关。结论:NF-κB可能参与了LN小鼠的发病机制,检测其在肾组织中的表达可作为反映狼疮肾组织活动病变的参考指标之一。     

Conversion of normal behavior to shiverer by myelin basic protein antisense cDNA in transgenic mice

Myelin basic proteins (MBPs) are coded by the single gene necessary for myelin formation in the central nervous system of the mouse. An antisense MBP mini-gene was constructed and used to determine the function of antisense DNA in transgenic mice. Several transgenic offspring of a founder transgenic mouse, AS100, were converted from the normal to mutant shiverer phenotype. Antisense MBP messenger RNA was expressed in these mice, and the endogenous MBP messenger RNA, the MBP, and the myelination in the central nervous system were reduced.     

Strain dependence of the antiproliferative action of interferon on murine erythroid precursors

Electrophoretically pure mouse interferon inhibits erythropoietin-dependent proliferation of committed erythroid precursors (CFU-E) obtained either from adult mouse bone marrow or from 14-day fetal mouse livers. The degree of inhibition is significantly influenced by the genotype of the cell donor; about ten times as much interferon is required to inhibit proliferation of CFU-E from C57BL/6 than is needed for comparable inhibition of CFU-E from BALB/c or Swiss mice. These strain-dependent results point to the existence of genes that influence the degree of the inhibitory effect of interferon on cell multiplication.