淮南猪品种遗传特异性的AFLP分析

试验利用25对引物对河南省地方品种淮南猪及引入品种杜洛克猪、长白猪、大约克猪4个品种猪进行AFLP分析。电泳检测结果发现,25对引物共检测到317条扩增片段,其中多态性片段79条,占24.92%。多态性统计分析表明,长白猪与大约克猪之间的遗传距离指数为0.0070,遗传关系最近;淮南猪与杜洛克猪、大约克猪遗传距离指数相近,分别为0.0073、0.0075,遗传关系较近;淮南猪与长白猪遗传距离指数为0.0078,遗传关系最远。结果显示,河南省地方品种淮南猪相对于其他引入品种特异性强,具有很高的育种价值,同时也证明了AFLP技术可以很好地检测猪种群内的遗传变异及猪种鉴别。     

用磁珠富集法从AFLP片段中分离鹅微卫星DNA标记

用磁珠富集法从AFLP片段中分离微卫星DNA标记，可以避免基因组文库的构建和筛选这一繁重的工作，既节省时间又节约费用。利用少量的内切酶组合建立一系列AFLP DNA，然后用生物素标记的简单重复序列（SSR）作探针与其杂交，杂交复合物固定到包被有链亲和素的磁珠上，经过一系列洗涤过程，含有SSR的AFLP片段被吸附到磁珠表面。这些片段经洗脱下来后，先用对应的AFLP引物扩增，再进行克隆和测序，根据SSR两端的保守序列设计引物，经过多态性分析后，便可得到微卫星DNA标记。用这种方法已成功地从6个中国地方品种鹅中分离到30个微卫星DNA标记。其中有11个引物表现为多态性好，特异性强且相互之间并不连锁。各品种平均多态信息含量（PIC）在0．513～0．546之间。说明这11个微卫星均为高度多态，可作为有效的遗传标记用于鹅品种间遗传多样性和系统发生关系的分析，并为建立鹅遗传图谱、重要经济性状的基因定位及标记辅助选择奠定基础。     

新疆6个地方绵羊品种遗传多样性的AFLP分析

本试验旨在从分子水平研究绵羊品种之间的遗传多样性,为科学评价新疆地方绵羊种质资源提供依据。利用筛选出的8对E+3/M+3引物,对6个地方绵羊品种基因组DNA进行AFLP扩增。根据AFLP分析结果,统计了每个引物组合在各品种中检测到的多态性条带和特异性条带,计算了6个绵羊品种的遗传相似系数和遗传距离,并构建了UPGMA聚类关系图。结果表明,8对引物组合共得到334条清晰可辨条带,平均每个引物组合产生41.75条多态性标记,多态性条带为103条,多态位点百分率为30.838%,同时各品种中还检测出特异性条带。通过UPGMA聚类分析,将6个地方绵羊品种种群划分为2类。因此,6个地方绵羊品种均得到较丰富的遗传多样性,其遗传相似系数和聚类结果与各绵羊品种的地理分布及现实状况相吻合。     

桑树多倍体育种材料遗传背景的AFLP分析

利用AFLP(Amplified Fragment Length Polymorphism)分子标记技术,即扩增片段长度多态性,从DNA分子水平对19份人工三倍体桑育种材料进行了遗传背景分析,构建了指纹图谱.19份育种材料可组配70个杂交组合,各材料间AFLP相似系数大于0.750 0的有54个组合,经AFLP分析认为,它们不宜作亲本间杂交组合;AFLP相似系数小于0.750 0的有16个组合,经AFLP分析认为它们适于作亲本间杂交组合.本研究从基因组DNA分子水平上为人工三倍体桑品种选育以及杂交亲本的选择和组配提供了遗传背景依据.     

桑树多倍体育种材料遗传背景的AFLP分析

利用AFLP(AmplifiedFragmentLengthPolymorphism)分子标记技术 ,即扩增片段长度多态性 ,从DNA分子水平对 19份人工三倍体桑育种材料进行了遗传背景分析 ,构建了指纹图谱。 19份育种材料可组配 70个杂交组合 ,各材料间AFLP相似系数大于 0 75 0 0的有 5 4个组合 ,经AFLP分析认为 ,它们不宜作亲本间杂交组合 ;AFLP相似系数小于 0 75 0 0的有 16个组合 ,经AFLP分析认为它们适于作亲本间杂交组合。本研究从基因组DNA分子水平上为人工三倍体桑品种选育以及杂交亲本的选择和组配提供了遗传背景依据     

地方鸡种与隐性白羽鸡遗传变异的AFLP指纹

利用6对AFLP引物组合对我国12个地方鸡种和引进鸡种隐性白羽鸡进行了遗传检测,统计了每个引物组合在各个品种中检测到的多态性条带和特异性条带,计算了13个鸡种的遗传相似系数和遗传距离,并据此构建了UPGMA聚类关系图,分析了所研究鸡种的遗传关系.结果表明:6对AFLP引物组合在13个鸡种中共检测到290条多态性条带,平均每个引物组合产生48.3奈多态性标记,同时在每个品种群体中还检测到了数量不等的特异性条带,其中寿光鸡和东乡黑鸡最多,为9条,旧院黑鸡、兴义矮脚鸡和隐性白羽鸡最少,为1条.13个鸡种聚为4类,其中隐性白羽鸡单独聚为一类,鸡种间的遗传相似系数及聚类结果与各个鸡种的地理分布、现实状况相吻合,从而表明AFLP指纹用于我国地方鸡种的遗传多态性分析、品种鉴定及品种间亲缘关系分析是可行的.     

哈萨克羊、阿勒泰羊、巴什拜羊遗传多样性的AFLP分析

利用11对AFLP引物组合,检测了哈萨克羊、阿勒泰羊、巴什拜羊池DNA遗传变异,构建了各品种的AFLP DNA 指纹图谱,根据AFLP分析结果,统计了每个引物组合在各品种中检测到的多态性条带,计算了3个品种的遗传相似系数,并据此构建了UPGMA聚类关系图,分析了它们的遗传关系.结果表明:11对AFLP引物组合在3个地方绵羊品种中共检测到310条带,平均每个引物组合产生28.18条带,变化范围在25～34条,其中多态性条带32条,占扩增总带教的10.32%,每对引物平均扩增多态性带2.909条.品种间的遗传相似系数以哈萨克绵羊与阿勒泰绵羊的最高(O.699),哈萨克绵羊与巴什拜绵羊最小(0.592),聚类结果与这些羊的育成史、分化及地理分布一致.     

中国6个地方猪种与3个外种猪氟烷基因PCR产物序列比较研究

本文应用PCR方法 ,体外扩增了中国地方猪和外种猪共 16个个体的氟烷基因 1814 9～ 1876 0位之间 6 12bp的片段 (包含完整的外显子 17和cDNAC1843 →T1843 突变位点 ) ,并进行了序列测定。结合网上下载的 5个中外猪种相同区段的序列进行了比较研究。结果表明 ,本文所测序列比网上公布的序列少 2bp ,在 1815 3,1815 4处有两胞嘧啶(C)缺失 ;皮特兰猪在cDNA的C1843 位点存在C1843 →T1843 突变 ;6 12bp的片段中共有 14个变异位点 ,皆为转换型突变 ,内含子 16中有 3个位点存在转换型杂合子 ;所有突变中 ,有 4处发生在外显子 17内 ,除香猪和皮特兰猪外均为同义突变 ;这些变异位点共形成 12种单倍型 ,其中扩增片段 5 14位 (全序列中为 186 6 2位 )的C→T突变为内江猪独有 ,构成了内江猪独特的单倍型 ,C18662 →T18662 突变具有品种特异性     

猪细小病毒PCR检测方法的建立与应用

根据已报道的猪细小病毒基因组序列，设计并合成了1对寡核苷酸引物，通过对影响PCR扩增因素的筛选，成功地从猪细小病毒感染的组织和细胞中扩增出445bp片段，回收该片段用EcoR I酶切得到了预期的结果，证实了该扩增片段的特异性。敏感性试验表明，该方法可以检测出10^-4个TCID50的病毒含量。利用该方法对临床上24份流产病科的检测，查出阳性16份，而同时利用HA检测阳性只有10份。这些结果说明本试验建立的PCR诊断方法灵敏度高、特异性强。     

中国地方猪种品种特异性遗传标签构建——以莱芜猪为例

我国拥有丰富的地方猪种资源,但地方猪种的保护和开发工作尚任重道远。建立完善可靠的生猪及猪肉产品追踪溯源技术体系对于建设优质安全猪肉品牌极其关键。传统的基于条形码、无线射频技术等的猪肉食品安全追溯技术,很难对从田间到餐桌的猪肉生产、屠宰、加工零售等环节实现全程精确而可靠的追溯。该研究以莱芜猪为例,对35个中外地方品种(中国地方猪种29个,培育品种2个,西方商品品种4个),共986头个体,进行60K芯片扫描判型;利用全基因组关联分析的case-control方法筛选出品种特异遗传位点,再结合主成分分析和邻近法进化树相互印证,构建了莱芜猪的品种特异性遗传标签组合并进行了验证:标签集为100个特异SNP位点,品种鉴别准确率达到100%。该研究所构建的品种特异性遗传标签可以准确地用于猪只个体及其猪肉产品的品种鉴别,为猪肉产品的追踪溯源奠定了坚实的技术基础,对我国地方猪种的品种保护和品牌猪肉深度开发提供了新的技术支撑。     

Molecular evidence for the infection of zoo chimpanzees by pig Ascaris

We here describe the transmission of the pig roundworm, Ascaris suum to chimpanzees maintained in the Copenhagen Zoo, Denmark. Using a technique for whole genome fingerprinting, amplified fragment length polymorphism (AFLP) and the technique PCR restricted fragment length polymorphism (PCR-RFLP) of the internal transcribed spacer region (ITS) of the nuclear ribosomal DNA, the worms from the chimpanzees were compared with Ascaris spp obtained from humans and pigs in order to identify the source of the infection. By the use of different distance and clustering based methods on the AFLP data set the worms from the chimpanzees were assigned to the same cluster as that of the worms from pigs. The PCR-RFLP analysis supported the AFLP results. Therefore, the zoo chimpanzees have required Ascaris infections by cross-infection from pigs. Pigs as a potential source of Ascaris infections for both captive and wild chimpanzees and other animals, therefore needs to be considered and appropriate steps taken to prevent such infection.     

分子标记在猪育种中的应用

本文较全面地介绍了RFLP，微卫星DNA，小卫星DNA，RAPD，AFLP以及单核苷酸多态性分子标记的研究进展及应用概况，并对分子标记在猪育种中的应用作了介绍。     

Molecular analysis of field strains of Mycoplasma capricolum subspecies capripneumoniae and Mycoplasma mycoides subspecies mycoides, small colony type isolated from goats in Tanzania.

A molecular analysis of strains of Mycoplasma capricolum subsp. capripneumoniae (M. capripneumoniae) and Mycoplasma mycoides subsp. mycoides, small colony type (M. mycoides SC) isolated from goats was performed using the amplified fragment length polymorphism (AFLP) and pulsed-field gel electrophoresis (PFGE) fingerprinting techniques. Among the 11 field strains of M. capripneumoniae from Tanzanian goats, two AFLP patterns were demonstrated, with 10 of the strains showing indistinguishable patterns. Five Kenyan strains of M. capripneumoniae produced three AFLP patterns, with two of them being indistinguishable from the 10 identical Tanzanian and one Ugandan strain (M74/93) isolated from sheep. The AFLP pattern of the type strain (F38(T)) was identical to two Kenyan strains (Baringo and G183/82). On PFGE analysis, all the examined M. capripneumoniae strains exhibited identical PFGE profiles.Five field strains of M. mycoides SC isolated from goats displayed identical AFLP patterns except for one strain which differed from others at only one position. The AFLP pattern of the type strain of M. mycoides SC (PG1(T)) was different from the field strains. The five field strains of M. mycoides SC produced identical PFGE profiles, which were, however, different from the type strain. The AFLP and PFGE profiles of M. mycoides SC strains from goats were identical to those of six strains isolated from cattle affected with contagious bovine pleuropneumonia (CBPP) in the same areas. The results of this study suggest a close epidemiological linkage between strains of M. capripneumoniae and between M. mycoides SC type, respectively, isolated from goats in Tanzania.     

Vergleich der histologischen Struktur der Kompakta der langen Röhren-knochen bei Maus, Hamster, Ratte, Meerschweinchen, Kaninchen, Katze und Hund während der Altersentwicklung

This paper reports on histological examinations of the diaphyseal structure of the long bones (humerus, radius, ulna, os femoris and tibia) of mouse, hamster, rat, guinea pig, rabbit, cat, and dog using six animals in three different age groups. There are considerable differences in structure between the species. The results show that with increasing height in the evolution of the species and period of life, as well as advancing age, the differentiation and complexity of bone structures increases. The differences in structure between the localization of bones within the individual species are not considerable. The species comparison results in great similarities between the bone structures of each of mouse and hamster, of rat and guinea pig, as well as of cat and dog. The bone structure of the animals examined becomes more similar to the human structure from mouse, hamster, rat, guinea pig, rabbit, and cat to dog.     

Genotyping of Mycoplasma gallisepticum and M. synoviae by Amplified Fragment Length Polymorphism (AFLP) analysis and digitalized Random Amplified Polymorphic DNA (RAPD) analysis

Mycoplasma gallisepticum (MG) and M. synoviae (MS) are the cause of considerable economic losses in the poultry industry. Molecular differentiation of avian Mycoplasma strains may be helpful in tracing infections and in the evaluation of implemented intervention strategies. Amplified Fragment Length Polymorphism (AFLP) has shown to be a powerful typing technique but the application for poultry Mycoplasma strains is very limited. The aim of this study was to evaluate the reproducibility and discriminatory power of AFLP HindIII/HhaI and AFLP BglII/Mfel for the inter- and intraspecies differentiation of avian mycoplasmas and to compare these test characteristics with digitalized Random Amplified Polymorphic DNA (RAPD) analysis. The reproducibility of RAPD, AFLP HindIII/HhaI and AFLP BglII/Mfel was 50-100, 97-98 and 86-99%, respectively. RAPD and both AFLP enzyme combinations were able to differentiate between five avian Mycoplasma species. For AFLP, five MG and four MS clusters could be identified. The phylogenetic tree for both enzyme combinations was comparable. For RAPD, four MG clusters could be identified. For MS, however, due to the poor reproducibility of the RAPD technique, no clear genogroups could be identified. On basis of the results of this study it can be concluded that AFLP is a powerful technique for the genotyping of avian mycoplasmas and that, although AFLP HindIII/HhaI generated patterns with less fragments, the final results showed homologous results.     

基于AFLP的燕麦遗传多样性研究

采用ＡＦＬＰ分子标记技术对来自中国、加拿大、澳大利亚和欧洲等国家的３４个有代表性的饲用燕麦品种及８个裸燕麦品种进行遗传多样性分析,从３０ 对ＥｃｏＲＩ／ＭｓｅＩ引物组合中筛选出５ 对多态性和清晰度较高的引物组合,共获得２６８条带,其中多态带１８５条,平均多态性检出率为６９．０％,Ｅ ＡＧＧ／Ｍ ＣＴＡ 的扩增效率最高达７８．６％。由Ｎｅｉ’ｓ指数（ｈ）估测,供试品种平均变异为０．１６６４,平均Ｓｈａｎｎｏｎ’ｓ信息指数估计值为０．２２０６;４２个燕麦品种间遗传相似系数为０．４８８１～０．９８８０,遗传距离的变化范围是０．０１２０～０．７１７２。通过ＡＦＬＰ 数据构建燕麦的ＵＰＧＭＡ 系统树,在遗传相似系数０．７４８水平,所有供试品种可聚为６类。皮燕麦与裸燕麦在遗传相似度０．５９处被明显分为２类,与形态学分类结果一致。由ＡＦＬＰ揭示的品种间遗传关系与品种实际来源基本一致,品种的遗传距离与其地理分布关系密切。     

Applicability of vital staining and tissue clearing to vascular anatomy and melanocytes' evaluation of temporal bone in six laboratory species

The purpose of the present study was to define the applicability of tissue clearing to the field of otology. We combined tissue clearing with vital staining perfusion via a pumping system to examine the vascular anatomy of temporal bones in laboratory animals. We used six different types of species including Korean wild mouse, mouse, Mongolian gerbil, hamsters and Guinea pigs. A mixture of Alcian blue reagent and 4% paraformaldehyde was circulated throughout the entire circulatory system of the animal via a perfusion pump system. Transparency images were obtained from the temporal bones according to the protocol of the SunHyun 3D Imaging Kit. In examining the inner surface of the tympanic membrane, flaccid part (pars flaccida) was positioned along the entire marginal area in Guinea pig. In the Guinea pig, unlike the other species, the cortical bone of the mastoid (bullae) was easily removed using cold instruments, allowing a direct approach to the enclosed structures. The distribution and pattern of cochlea melanocytes were compared among the species. “Mobius strip”‐like accumulated melanocytes in vestibules were shown in both the Korean wild mouse and mouse. The collateral blood supply to the cochlea in six different species was checked in various pattern. Combining dye infusion with tissue‐clearing techniques, we documented the middle ear and transparent inner ear structures in six different species. The information and associated images will help other researchers to develop hypotheses and design experimental investigations.