Southern杂交方法用于近交系小鼠H-2基因检测

目的建立一种简便、快捷、准确的检测方法用于近交系小鼠的遗传检测。方法根据近交系小鼠的H-2基因序列设计相应的探针,并标记生物素,利用微孔板Southern杂交技术,使探针与模板DNA杂交,再加入亲和素标记的辣根过氧化物酶进行酶显色反应,通过酶标仪检测杂交结果,以确定近交系小鼠的基因型。结果57BL／6和C57B：／10为H-2^b型;DBA／2和Scid为H-2^d型;615和C3H为H-2^k型;NCPC／2、TA1、TA2和T739均为H-2^b型。结论通过Southern杂交检测可以确定近交系小鼠的基因型。该检测方法简便、易行,检测结果客观,可以应用于近交系小鼠的遗传检测。     

Establishment and characterization of the Komeda diabetes-prone rat as a segregating inbred strain.

The Komeda diabetes-prone (KDP) rat is a spontaneous animal model of human autoimmune type 1 diabetes. By positional cloning of the non-MHC major susceptibility locus lddm/kdp1, we recently identified a nonsense mutation in Cblb and also found that lymphocytes of KDP rats infiltrate into various tissues, indicating autoimmunity. The maintenance and production of KDP rats has been a critical problem owing to the poor reproductive ability of diabetic animals. To solve the problem, we here established the KDP rat as a segregating inbred strain. We first identified animals that were heterozygous at the lddm/kdp1 region in a breeding colony of KDP rats. The heterozygous region spans at least from D11Yok1 to Cblb on rat chromosome 11. By mating between the heterozygous rats, we obtained homozygotes, heterozygotes and wild-types with the expected ratio of 1:2:1 and found that only the homozygotes developed diabetes, suggesting that these genotypes represent those of lddm/kdp1. We then tried to maintain KDP rats by mating between the heterozygotes, which resulted in a segregating inbred strain. Within 210 d of age, about 80% of lddm/kdp1 homozygotes developed diabetes with severe insulitis, while neither heterozygotes nor wild-types developed diabetes. The phenotypic characteristics of the homozygotes are the same as those of progeny of diabetic parents in the original KDP rats. The segregating inbred KDP rat strain described here would serve as a useful animal model for autoimmune diseases, including type 1 diabetes.     

山医群体近交系中国地鼠(Cricetulus griseus)的研究——Ⅰ、驯养与繁殖

经过12年工作,将野生中国地鼠驯养和近亲交配,繁育成山医群体,其中A-30家系已繁殖到22代。经遗传监测,证明20代鼠已育成近交系。文中对饲养条件、繁殖方法、近交系的建立等进行了讨论。     

DA大鼠的繁殖性能及部分生物学特性参数的测定

目的建立我国新的近交系品系DA种群,观察其生长繁殖性能及部分生物学特性。方法从丹麦M＆B公司引进近交系大鼠DA,建立种群,从其扩大群中随机抽取15对鼠,统计其繁殖率;抽取50只DA鼠（雌雄各半）测其部分血液生化。结果近交系种群扩大成功,DA大鼠平均产仔数为5．5,成活率为90.8％,所测生理生化值接近Wistar大鼠。结论DA大鼠已适应我国环境,种群引进成功;性别对脏器重量、系数和血生化影响显著。     

NIH/Ola: a highly productive inbred strain of laboratory mouse

According to historical records the NIH/Ola strain was developed from outbred 'Swiss' mice imported into the USA by Dr C. Lynch in 1926. A comparison of biochemical markers in strain NIH/Ola and other strains such as SJL and SWR derived from the same foundation stock supports the historical records. Data on litter size, length of gestation, bodyweight to 70 days of age, and reproductive performance when housed in polygamous groups of up to 10 females per male were compared with similar data on inbred CBA/CaOla and C57BL/10ScSnOla mice. NIH/Ola inbred mice had an exceptional reproductive performance, producing about 1.8 young weaned/female/week when housed as monogamous pairs over a period of 20 weeks, compared with less than 1.0 young/female/week with the other 2 strains. NIH/Ola mice were also extremely tolerant to mating in polygamous groups of up to about 8 females per male. Mating ratios of over 2-3 females per male resulted in a marked decline in the total number of litters produced per female in C57BL/10ScSnOla and CBA/CaOla, but this reduction was not nearly so marked in NIH/Ola. It is concluded that the NIH/Ola inbred strain may be particularly useful in studies such as teratology where a high reproductive performance needs to be combined with the advantages of a fully inbred strain.     

IRM-2近交系小鼠的生殖生长特性

目的获取IRM-2近交系小鼠的生殖生长特性的有关资料.方法ICR/JCL为母本,以615小鼠为父本杂交选育的近交系小鼠IRM-2,现已繁育至第38代.经过对其生殖、生长特性的观察和测定来取得相关资料.结果该小鼠出生后45d性成熟;繁殖能力强,平均每窝产仔数为8只以上,最高可达15只;生长发育迅速,出生后60d体重可达28g以上,各脏器重均高于亲代鼠.结论从各项指标来看,IRM-2小鼠是优良的近交品系小鼠.     

A genetic quality testing system for early stage embryos in the mouse.

We have established a genetic quality testing system for early stage embryos of the mouse. A method of preparation of template DNA for PCR was established using the lysis buffer (1 x PCR reaction buffer supplemented with proteinase K at a concentration of 40 microg/ml) developed by the authors. We demonstrated that two 8-cell embryos of an inbred strain provide sufficient volumes of template DNA for PCR to identify the strain of embryos using four microsatellite markers (D3Mit54, D5Mit18, D6Mit15 and D8Mit50) differentiating 13 inbred strains of mice. This system will be useful in embryo banks that have recently been established worldwide for demonstrating the genetic accuracy of a given strain prior to recovery of live animals.     

Heligmosomoides polygyrus (=Nematospiroides dubius): the development of self-cure and/or protection in several strains of mice.

Strains of outbred (ICR/CD1 and S--W) and inbred (BALB/C and C57BL/6) mice vaccinated subcutaneously (SQ) with 500, 1,000, or 2,000 exsheathed Heligmosomoides polygyrus larvae developed varying levels of protection upon subsequent oral challenge with larvae. In contrast, the inbred C3H/HEJ strain failed to develop protection at any dosage level tested. ICR/CD1 mice vaccinated intraperitoneally with exsheathed larvae developed a high level of resistance but exhibited extensive adhesions of the viscera. When ensheathed larvae were used for vaccination, ICR/CD1 mice developed a moderate level of protection; but 1% of the vaccine dose was recovered in the intestine as adult stages. Both the inbred and outbred strains given multiple oral infections developed a protection response similar to that strain's response following parenteral vaccination. The specificity of this protection was demonstrated using various complex foreign antigens. In contrast, the self-cure response was observed only in the S--W strain.     

山医群体近交系中国地鼠血液生理生化指标的测定分析

目的对山医群体中国地鼠近交A、E家系主要血液生理、生化和血清电解质指标进行检测分析。方法对山医群体中国地鼠近交A、E家系成年动物进行空腹采血,常规方法测定13项血液生理、12项生化以及4项电解质指标,分别对相同性别不同家系间和同一家系不同性别间指标进行统计分析。结果相同性别A、E家系间动物血小板（PLT）、中间细胞数（MID）、谷丙转氨酶（ALT）、总胆固醇（TC）、总胆红素（TBiL）、肌酐（Cr）和血钾（K）差异有显著性;相同家系雌、雄动物间PLT、TBiL和尿酸（UA）差异有显著性。结论 A、E家系动物间产生了一定的血液生理、生化性状的分离。     

中国地鼠山医群体近交系E繁殖性状的遗传力估计

目的对中国地鼠山医群体近交系E的一些繁殖性状进行遗传力估计分析。方法采用平均信息约束最大似然法（AIREML）,利用WOMBAT软件进行处理。结果产仔数、离乳数、胎间隔2-1和胎间隔3-2的遗传力均较低,分别为0.05、0.096、0.182和0.116。结论中国地鼠山医群体近交系E繁殖性状的遗传力均属于低遗传力。     

家蚕近交系遗传纯度的RAPD检测

目的分析家蚕近交系IS-c108A的遗传纯度,为家蚕实验动物化的培育工作提供指导。方法应用经过筛选的20条随机引物对家蚕近交系IS-c108A(F10)的3个蛾区各30个个体和该近交系的亲本系统c108、对照实用化品种871各30个个体的基因组DNA进行RAPD扩增,计算个体间和蛾区间的相似系数及遗传距离。结果家蚕近交系IS-c108A(F10)的3个蛾区内的多态性带频率分别为1.807%、1.841%、1.841%,平均为1.830%;起点亲本c108个体间多态性带频率为7.207%,对照品种871个体间的多态性带频率为7.08%;而近交系IS-c108A与c108之间的多态性带频率为49.20%,c108和871品种之间的多态性带频率为58.33%。家蚕近交系IS-c108A10的3个蛾区内个体之间遗传相似系数的平均值分别为0.99581、0.99555、0.99551,总平均为0.99562。结论家蚕近交系IS-c108A(F10)已具有较高的遗传纯合度,家蚕具有易于获得高纯的有利条件。     

近交系剑尾鱼的遗传生化标记研究

目的通过研究对RR-B、RW-H、BY-F三个近交系和非选育剑尾鱼的研究比较,建立近交系剑尾鱼的遗传生化标记检测技术。方法依照国标GB／T14927.1-2008的遗传操作规程优化实验条件,对三个近交品系及非选育群剑尾鱼的不同组织的6个遗传生化位点进行研究,以得到其遗传生化图谱。结果在6个生化位点中,同一品系剑尾鱼同工酶存在组织特异性,多数同工酶在肝中活性较强。同一生化位点在不同品系间存在差异。RR-B系在葡萄糖磷酸异构酶（Gpi）、6-磷酸葡萄糖脱氢酶（Gpd）位点表现出特异性条带,RW-H系在过氧化氢酶（Ce）位点表现出特异性条带。同一生化位点在各品系内表现较为一致,而在非选育剑尾鱼中在上述三个生化位点表现出多态性。在酯酶（ES）、碱性磷酸酶（AKP）、乳酸脱氢酶（LDH）位点,各品系谱带不易区分其差异。结论建立了剑尾鱼近交系生化标记检测技术,可望用于近交系剑尾鱼的遗传质量监测。     

Simulation of the distribution of parental strains’ genomes in RC strains of mice

Recombinant Congenic strains (RC strains) were developed to facilitate mapping of genes influencing complex traits controlled by multiple genes. They were produced by inbreeding of the progeny derived from a second backcross from a common `donor' inbred strain to a common `background' inbred strain. Each RC strain contains a random subset of approximately 12.5% of genes from the donor strain and 87.5% of genes from the background strain. In this way the genetic control of a complex disease may be dissected into its individual components. We simulated the production of the RC strains to study to what extent they have to be characterized in order to obtain sufficient information about the distribution of the parental strains' genomes in these strains and to acquire insight into parameters influencing their effectiveness in mapping quantitative trait loci (QTLs). The donor strain genome in the RC strains is fragmented into many segments. Genetic characterization of these strains with one polymorphic marker per 3.3 centiMorgans (cM) is needed to detect 95% of the donor strain genome. The probability of a donor strain segment being located entirely in between two markers of background strain origin that are 3 cM apart (and hence escaping detection) is 0.003. Although the donor strain genome in the RC strains is split into many segments, the largest part still occurs in relatively long stretches that are mostly concentrated in fewer than 13 autosomes, the median being 9 autosomes. Thus, in mapping QTLs, the use of RC strains facilitates the detection of linkage. Received: 20 December 1996 / Accepted: 23 July 1997     

野生来源TW小鼠近交系培育及其生物学特性的研究

目的　培育中国特有TW (TianjinWild)野生小鼠来源的实验近交小鼠品系 ,筛选其重要的生物学特征 ,丰富现有实验动物基因库。方法　严格按照国际培育近交系动物标准 ,即每代繁育的雌雄小鼠严格按照同胎雌雄 (兄妹 )进行交配。结果　经过 1 2年的精心培育 ,培育TW小鼠至F2 2代 ,其他 1 5地野生小鼠品系均因故中断。结论　该项研究将为丰富国际现有实验小鼠基因库资源 ,建立抗肺肿瘤特性实验小鼠模型动物奠定坚实基础。     

Quiet mutations in inbred strains of mice

The year 2009 is the 100th anniversary of the founding of the first inbred strain of mouse, called DBA. During the last 100 years, inbred strains have proved their value for biomedical research and the number of such strains has mushroomed to over 450, each with different genotypic and phenotypic characteristics and useful for the study of disease and normal function. However, although inbred strains are stable, they are not fixed entities and researchers need to be aware of the phenomena of new mutations and of genetic drift, which occur within all mouse colonies. If the mutations are what we term in this review 'quiet mutations', then they might result in rather unexpected and sometimes tremendously valuable results. Here, we discuss these phenomena and look at how new genomic technologies might help us to detect 'quiet mutations' and use them to our advantage.     

Different degree of ileal colonization by segmented, filamentous bacteria in two strains of mice.

Segmented, filamentous bacteria (SFBs) are autochthonous, apathogenic inhabitants of the ileum of various animal species. Outbred Swiss (Cpb:SE) mice have significantly higher degrees of SFB colonization than do inbred BALB/c mice. The present studies were carried out to identify determinants of this strain difference. In a cross-fostering experiment it was shown that SFB colonization of the pups is determined by the strain of the pups themselves rather than by the strain of the nursing dam. Thus, maternal effects may not be involved in SFB colonization. In a cross-infecting experiment using germ-free and SFB-positive animals of the two mouse strains, it was found that ileal SFB colonization is determined by host characteristics rather than by origin of the SFBs. Thus, SFBs that are specific for a given mouse strain may not exist in the two strains of mice. It is concluded that the mouse strain difference in SFB colonization is determined by host characteristics, which probably have a genetic basis.     

红鲫C1HD近交系RAPD标记的建立

目的建立红鲫C1HD近交系的RAPD标记。方法从80条随机引物中筛选出20条扩增效果和多态性较好的引物,对8尾红鲫C1HD近交系和8尾普通红鲫基因组DNA进行RAPD扩增。结果S333引物扩增出一条特异性条带,大小约为2.1 kb。结论S333引物扩增出的特异性条带可以作为区分普通红鲫与红鲫C1HD近交系的分子遗传标记。     

Linkage of Nat and Es-1 in the mouse and development of strains congenic for N-acetyltransferase

The human polymorphism in the hepatic enzyme N-acetyltransferase (NAT) affects the rate at which individuals acetylate, and in many cases detoxify, aromatic amine and hydrazine drugs and xenobiotics. Differences in NAT activity are known to affect individual susceptibility to drug toxicities and are thought to play a part in some spontaneous disorders. A mouse model for the human acetylation polymorphism has been previously characterized and involves the A/J (slow acetylator) and C57BL/6J (rapid acetylator) inbred strains. Strain distribution analysis of 40 A x B and B x A recombinant inbred (RI) strains indicated linkage between the N-acetyltransferase gene (Nat) and the esterase 1 (Es-1) gene, located on mouse chromosome 8. A double backcross involving 107 animals confirmed the recombination frequency between Nat and Es-1 to be 12 +/- 3% (mean +/- SE). The information obtained in the backcross and RI studies was combined, yielding a 13 +/- 2.8% (mean +/- SD) recombination frequency. The Es-1 genotype was determined in our newly developed congenic strains A.B6-Natr and B6.A-Nats. The B6.A-Nats strain has the Es-1 genotype of its inbred partner, the B6 strain, and the A.B6-Natr strain has the Es-1 genotype of the donor strain. These congenic strains will be important in determining the role of the NAT genotype in susceptibility to arylamine-induced cancer and other disorders.     

Mouse phenome research: implications of genetic background

Now that sequencing of the mouse genome has been completed, the function of each gene remains to be elucidated through phenotypic analysis. The "genetic background" (in which each gene functions) is defined as the genotype of all other related genes that may interact with the gene of interest, and therefore potentially influences the specific phenotype. To understand the nature and importance of genetic background on phenotypic expression of specific genes, it is necessary to know the origin and evolutionary history of the laboratory mouse genome. Molecular analysis has indicated that the fancy mice of Japan and Europe contributed significantly to the origin of today's laboratory mice. The genetic background of present-day laboratory mice varies by mouse strain, but is mainly derived from the European domesticus subspecies group and to a lesser degree from Asian mice, probably Japanese fancy mice, which belong to the musculus subspecies group. Inbred laboratory mouse strains are genetically uniform due to extensive inbreeding, and they have greatly contributed to the genetic analysis of many Mendelian traits. Meanwhile, for a variety of practical reasons, many transgenic and targeted mutant mice have been created in mice of mixed genetic backgrounds to elucidate the function of the genes, although efforts have been made to create inbred transgenic mice and targeted mutant mice with coisogenic embryonic stem cell lines. Inbred mouse strains have provided uniform genetic background for accurate evaluation of specific genes phenotypes, thus eliminating the phenotypic variations caused by mixed genetic backgrounds. However, the process of inbreeding and selection of various inbred strain characteristics has resulted in inadvertent selection of other undesirable genetic characteristics and mutations that may influence the genotype and preclude effective phenotypic analysis. Because many of the common inbred mouse stains have been established from relatively small gene pools, common inbred strains have limitations in their genetic polymorphisms and phenotypic variations. Wild-derived mouse strains can complement deficiencies of common inbred mouse strains, providing novel allelic variants and phenotypes. Although wild-derived strains are not as tame as the common laboratory strains, their genetic characteristics are attractive for the future study of gene function.