鸡的小卫星探针产生的猪DNA指纹图

以一个鸡的小卫星cMS18为探针,对五指山小型猪、长白猪及枫泾猪进行了DNA指纹分析。结果表明,五指山小型猪的相似系数(0.721)显著高于长白猪(0.503)和枫泾猪(0.484),在遗传背景上也与后两者相距甚远。家系分析表明, 鸡的小卫星探针产生的猪的DNA指纹图的遗传符合孟德尔规律。 Abstract:A chicken minisatellite probe cMS18 was used to generate DNA fingerprints of Wuzhisshan microping,Landrace pig and Fengjing pig.It was revealed that similarity coefficient of Wuzhishan microping(0.721)was much higher than both that of Landrace pig(0.503)and of Fengjing pig(0.484).It was shown that Wuzhishan micropig was genetically distant to Landrace pig and Fengjing pig.Analysis of a porcine pedigree using cMS18 probe manifested that the segregation of DNA fingerprint bands was consistent with the Mendelian fashion.     

枫泾猪、香猪和长白猪的DNA指纹图分析

本文用M13和3个人源小卫星探针(33.6、 33.15、α-珠蛋白-3’HVR)获得了枫泾猪、香猪及长白猪的DNA指纹图,分析了各品种内和品种间的遗传变异性。结果表明,各品种的平均DNA指纹图相似系数在0.440-0.532的范围内,极显著地高于品种间的相似系数(0.223-0.307)。各品种的DNA指纹图还存在着一些品种特异性图带,这些图带在以后的基因定位中值得重视。 Abstract:This paper demonstrated DNA fingerprints of Fengjing,Xiang and Large White breeds of swine generated by M13 and three human minisatellite probes(33.6,33.15 andα-globin-3’HVR),and determined the intrabreed and interbreed variabilities of swine DNA fingerprints.It was shown that the mean band sharing probability within breeds was 0.440-0.532,significantly higher than that between breeds(0.223-0.307).Some breed-specific bands were noted in each breed,which should draw our attention in the course of gene mapping.     

应用多重PCR和基因扫描技术对五指山猪13个家系32个微卫星基因座的遗传分析

五指山猪是我国著名的珍稀畜种,体型小、性成熟早、遗传稳定、近交程度高,可用于医学、药学和生物材料等方面。作者利用多重PCR和基因扫描技术对五指山猪13个家系的32个微卫星基因座进行遗传检测。统计各家系的等位基因组成,计算各家系的平均杂合度和多态信息含量(PIC);结果,各家系32基因座的平均等位基因数为13.66个,平均PIC为0.731,平均杂合度为0.559。表明海南五指山猪具有丰富的遗传多样性。这些结果对于海南五指山猪的保种、定向选育、开发利用都具有重要的指导意义。     

用DNA指纹法研究野生天鹅种群的遗传分化

DNA指纹技术自问世以来,已被广泛地用于法医学上个体识别和亲缘关系的鉴定。此外,人源DNA指纹探针(即多位点型小卫星探针)还被用于动植物等的研究,特别是在研究动物的繁育规律方面成效显著。近一两年来,DNA指纹技术也在分子水平上用来研究种群分化,为物种起源研究开辟了新的途径。本文利用人源多位点型小卫星探针,对分布在英国和美国的四种天鹅(小天鹅、疣鼻天鹅、大天鹅和喇叭天鹅)的种群结构和天鹅小卫星的分化初步进行了研究。用酶HaeIII和探针pSPT19.6分析了分布在英国的疣鼻天鹅种的两个群体(洛锡安群体和阿伯茨伯里群体)的DNA指纹图,结果表明两群体之间的遗传变异性大于群体内的遗传变异性;洛锡安群体内小卫星的变异程度较高。虽然小卫星在基因组中的作用尚不清楚,但某些天鹅小卫星在种间发生了分化,因而同一个种内不同个体的DNA指纹图有一些共同的特征,导致DNA指纹图的物种特异性。DNA指纹分析表明:小天鹅、大天鹅和喇叭天鹅在遗传上比较接近,而疣鼻天鹅与它们在遗传上相距较远,这与原来对天鹅种下分类情况是一致的。就遗传变异程度而言,在英国仕林布里奇的小天鹅为最高,英国卡拉乌尔洛克的大天鹅次之,英国阿伯茨伯里的疣鼻天鹅居第三,美国蒙大拿的喇叭天鹅最低。我们认为,基因迁移和遗传漂变对     

罗伯逊易位群体遗传结构的DNA指纹分析

用微卫星探针（GGAT）4得到了罗伯逊易位猪群体内共9个家猪品种的DNA指纹图。1－9品种依次为：13／17罗伯逊易位纯合子猪群体、13／17罗伯逊易位杂合子猪群体、13／17罗伯逊易位杂合子猪互交所得到的正常核型猪群体（杂交0号）、丹系长白猪群体、加系双肌臂杜洛克猪群体、加系双肌臀大约克猪群体、加系双肌臀长白猪群体、丹系长白猪与13／17易位杂合子猪产生的杂交后备猪群体（杂交1号）、杜洛克猪与13／17易位杂合子猪产生的杂交后备猪群体（杂交2号）。根据指纹带型计算了每个群体的相似系数（F）、平均等位基因频率（Q）、最底平均杂合率（H）和遗传距离指数（D）。9个品种的相似系数介于0．546－0．931,根据各品种之间的遗传距离指数作出矩阵图,并据此绘制出品种间亲缘关系树状聚类图。     

四个探针产生的家禽 DNA 指纹图谱

对四个多位点小卫星探针 33.6,33.15,α珠蛋白-3’HVR 和 M13用于家禽的DNA 指纹分析的可行性进行了探讨,较详细地报导了用不同探针在鸡、鸭、鹌鹑上产生 DNA 指纹图的方法.结果表明,用我们所采用的方法,四个探针都能在鸡、鸭、鹌鹑上产生信息量大、分辨率较高的 DNA 指纹图.     

海南五指山猪遗传多样性的微卫星分析

海南五指山猪(Wuzhishan pig)是我国珍稀的小型猪品种。它的解剖学、生理学、疾病发生机理等都与人类极其相似,具有重要的应用价值,可用于心血管病、皮肤烧伤和新药评价等许多方面的研究。作者利用多重PCR和基因扫描技术对五指山猪32个微卫星基因座进行了遗传检测。统计了全群、核心群和近交群的等位基因组成,并计算了它们的平均纯合率、多态信息含量(PIC)和平均杂合度。结果显示,全群、核心群和近交群32个基因座的平均等位基因数分别为14.590个、9.410个、13.660个,平均基因纯合率分别为44%、44.3%、44%,平均PIC分别为0.742、0.708、0.731,平均杂合度分别为0.560、0.558、0.559。这表明海南五指山猪虽然遗传基础比较稳定,但仍具有较大的遗传变异。上述结果对于海南五指山猪的保种、定向选育、开发利用都具有重要的指导意义。     

五指山猪和长白猪背最长肌差异基因的筛选与注释

为了筛选五指山猪和长白猪背最长肌组织差异表达基因,本研究采用RNA-seq技术和生物信息学方法对6月龄、8月龄五指山猪和长白猪的背最长肌进行转录组测序分析,并对差异表达基因进行GO和KEGG Pathway显著性富集分析。通过与猪基因组比对分析后发现,在6月龄和8月龄五指山猪和长白猪均有约44 000 000条Clean reads能够比对到猪基因组上相关基因,每个文库均获得约18 200个表达基因,而转录本的数据为25 000个。在6月龄五指山猪与长白猪中共有30个基因差异表达,在8月龄五指山猪与长白猪中获得29个差异表达的基因。这些差异表达的基因主要富集在与糖酵解代谢、MAPK信号代谢通路和胰岛素信号通路等肌肉发育信号通路,通过筛选获得了与肌肉生长发育有关的候选基因,为探索五指山猪肌肉发育的分子机制提供了理论依据。     

13/17染色体易位纯合子猪DNA指纹分析

13/17染色体易位纯合子猪是孙金海等于1991年通过易位杂合子互交在国内外首次获得的一种新遗传类型家猪[2n=36,XY或XXrob(13;17)]。本研究用噬菌体M13mp18单链小卫星DNA作探针对该种新遗传类型猪、13/17染色体易位杂合子猪及正常核型家猪进行了DNA指纹分析。采用限制性内切酶HinfⅠ和HaeⅢ均成功地获得高分辨的、具有个体特异性的DNA指纹图谱。研究结果表明,不同内     

纵纹腹小鸮DNA指纹谱探针的筛选和初步应用

以人工合成的微卫星序列 (GTG) 5,(GT) 8,(CAC) 5和人源小卫星 33 1 5作引物 ,扩增纵纹腹小的基因组DNA ,产生多态性DNA片段 ,回收了 8个表现个体特异性的片段。当用小的基因组总DNA探针与它们杂交时 ,其中 2个表现阳性 ,说明PCR方法扩增出的高变异产物含有重复序列。用含重复序列的个体特异性PCR产物作探针 ,与无关个体小基因组DNA的HaeⅢ酶切产物进行DNA印迹 ,获得了变异性较高的DNA指纹图谱。且通过对京白鸡家系分析表明 ,用小基因组DNA的PCR产物分离制备的探针所获得的DNA指纹图带能够稳定的遗传。因此 ,高变异的PCR产物可以有效地用作DNA指纹探针。     

白色基因座(I)在中国地方猪种毛色遗传中的作用研究

通过利用PCR—RFLP和PCR—SSCP技术对中国地方猪种KIT基因内含子17、18的序列进行多态性分析。结果表明：内含子17上的替换突变(G→A)发生于毛色为白色的个体——白色五指山猪、大白猪、长白猪上,其基因型(AB型)频率分别为1、1和0．8;其他中国地方猪种的此基因型频率均为0。内含子18上的缺失突变(AGTT)也同样发生在上述3个猪种的白色个体中,其基因型(AA型)频率分别为1、1和0．93;而且同样在其他的地方品种中其基因型频率均为0。这充分证明KIT基因对于猪的白毛色有重要的调控作用,而且I基因座对于其他的经典遗传基因座有上位作用。另一方面,中国地方猪种荣昌猪虽然在表型上与引入猪种大白猪、长白猪相似(白毛色),但是在KIT基因上发生的突变完全不同,推测它们分别属于不同的毛色遗传体系。     

利用微卫星标记分析七品种（类群）小型猪的遗传多样性

采用世界粮农组织（FAO）和国际动物遗传学会（ISAG）推荐的27个微卫星,对久仰香猪、剑白香猪、从江香猪、环江香猪、黑香猪（贵州省种猪场）、五指山猪和滇南小耳猪等我国7小型猪种（类群）及杜洛克、长白和大白等3个外来猪种的群内遗传变异性和群间遗传差异进行了分析。结果表明,7个小型猪品种（类群）均有较高的群内遗传变异,但久仰、剑白、从江和环江4个香猪类群的群内遗传变异（平均多态信息含量（PIC）为0.61～0.64）显著低于其他3个小型猪种（平均PIC为0.80～0.84）。久仰、剑白、从江和环江4个香猪类群彼此间的遗传差异较小（奈氏标准遗传距离为0.12~0.22）,但它们与其他3个小型猪种有较大的遗传差异（奈氏标准遗传距离为1.61～1.96）,与3个外来猪种的遗传差异更大（奈氏标准遗传距离为1.99～3.30）。通过聚类分析,可将这些猪种清晰地分为3大类,久仰、剑白、从江和环江4个香猪类群紧密地聚为一类,其他3个小型猪种聚为一类,3个外来猪种聚为另一类。这一结果基本符合这些猪种的地理分布和品种来源。
     

五指山猪IGF2基因5′调控区单核苷酸多态性分析

利用PCR产物直接测序法, 对五指山猪、滇南小耳猪、香猪、梅山猪和大白猪共60个样本的IGF2基因5＇调控区部分片段的单核苷酸多态性进行了研究。找到13个SNP, 分别是: C5872T、C5888T、A5976G、C6010T、T6029A、C6037T、C6043T、C6063T、C6112T、C6164T、G13520A、G13563A和G13669A。T6029A为T←→A碱基颠换, A5976G、G13520A、G13563A和G13669A为A←→G转换, 其他均为C←→T转换。针对13个SNP位点得到23种组合基因型。统计各位点等位基因和基因型以及各组合基因型在总群体与各品种内的分布频率, 发现3个小型猪在A5976G、C6164T和G13669A位点上的优势等位基因均分别为G、T和A, 而梅山猪和大白猪的优势等位基因均分别为A、C和G; H19型为3个小型猪的特征组合基因型, 而另两个猪品种为H15型。同时对123头五指山猪IGF2基因C5888T位点进行了PCR-RFLP分析, 研究表明该位点C为优势等位基因(0.8536), CC为优势基因型(0.7235)。卡方检验表明该位点处于Hardy-Weinberg平衡状态。这些结果可为五指山猪等小型猪的生长发育规律、矮小机制等方面的研究提供遗传学依据。     

中外11个猪种A-FABP基因微卫星遗传变异的研究

利用微卫星技术对五指山猪、沂蒙黑猪、汉江黑猪、莱芜猪、北京黑猪、民猪、成华猪、内江猪、二花脸猪、巴马香猪和大白猪11个猪种共420头猪的脂肪细胞脂肪酸结合蛋白基因内含子的微卫星序列的遗传变异进行了研究。研究表明：（1）中国猪种在A-FABP基因微卫星基因座上有极高的的多态性,五指山猪的PIC最高（PIC=0.7904）并检测到11个等位基因,但北京黑猪仅检测到两个等位基因。（2）本实验中,只有北京黑猪、民猪、巴马香猪和大白猪A-FABP基因微卫星在群体中的分布达到平衡状态。（3）群体间的基因分化系数分析表明,中国猪种在该微卫星基因座上分化程度平均约为40.83%。Abstract：The genetic variations of microsatellites in intron 2 of the porcine adipocyte fatty acid-binding protein (A-FABP) genes were investigated in 420 pigs including Wuzhishan pig,Yimeng black pig,Hanjiang black pig,Laiwu pig,Beijing black pig,Min pig,Chenghua pig,Neijiang pig,Erhualian pig,Bama xiang pig and Large White pig.The results suggested as follows：（i）PIC of the Wuzhishan pig breed is the highest（0.7904） and 11 alleles were detected. Compared with Large White pig,Chinese pig breeds showed a great polymorphism of A-FABP microsatellites except Beijing black pig in which only 2 alleles were detected.（ii）Only Min pig, Bama xiang pig, Beijing black pig and Large White pig were in Hardy-Weinberg equilibrium. （iii）The analysis of genetic differentiation showed that the average value of the A-FABP gene differentiation of 10 Chinese pigs is about 40.83%.     

Genetic diversity and population structure of six Chinese indigenous pig breeds in the Taihu Lake region revealed by sequencing data

The Chinese indigenous pig breeds in the Taihu Lake region are the most prolific pig breeds in the world. In this study, we investigated the genetic diversity and population structure of six breeds, including Meishan, Erhualian, Mi, Fengjing, Shawutou and Jiaxing Black, in this region using whole‐genome SNP data. A high SNP with proportions of polymorphic markers ranging from 0.925 to 0.995 was exhibited by the Chinese indigenous pigs in the Taihu Lake region. The allelic richness and expected heterozygosity also were calculated and indicated that the genetic diversity of the Meishan breed was the greatest, whereas that of the Fengjing breed was the lowest. The genetic differentiation, as indicated by the fixation index, exhibited an overall mean of 0.149. Both neighbor‐joining tree and principal components analysis were able to distinguish the breeds from each other, but structure analysis indicated that the Mi and Erhualian breeds exhibited similar major signals of admixture. With this genome‐wide comprehensive survey of the genetic diversity and population structure of the indigenous Chinese pigs in the Taihu Lake region, we confirmed the rationality of the current breed classification of the pigs in this region.     

湖北省三品种猪27个微卫星座位的遗传变异

采用国际动物遗传学会（ISAG）和联合国粮农组织（FAO）共同推荐的27个微卫星标记,对湖北省3个主要地方猪种（通城猪、清平猪和阳新猪）的遗传变异进行了检测。计算出各个品种的基因杂合度、各个座位的多态信息含量及品种间的遗传距离。结果表明,3个地方猪种的平均基因杂合度分别为0.7489、0.6987和0.6273,遗传多样性比较丰富;通城猪和清平猪亲缘关系较近,而两者与阳新猪亲缘关系略远。     

Comparison of the semen characteristics of Fengjing, Meishan and Yorkshire boars

Semen characteristics of Chinese Fengjing, CHinese Meishan and American Yorkshire boars were examined. Samples were collected from 24 boars: 6 Fengjing, 12 Meishan and 6 Yorkshire. Three semen characteristics and 6 biochemical evaluations of semen or seminal plasma were analyzed. The whole semen parameters measured were gelatinous (gel) and gel-free volume and progressive motility. Fengjing boars were higher in gel volume than Meishan (P < 0.05) but not Yorkshire boars (P > 0.25), while Yorkshire boars were higher than Meishan boars in gel volume (P < 0.10). The gel-free volume was higher in Yorkshire and Fengjing boars than Meishan boars (P < 0.10), but Fengjing gel-free volumes did not differ from Yorkshire gel-free volumes (P > 0.80). However, the only difference detected for progressive motility was between Fengjing and Meishan boars (78.5 vs 74.5%; P < 0.10). Sperm concentration was higher in Meishan than Yorkshire boars (P < 0.01) although these breeds did not differ from Fengjing boars (P > 0.18). The biochemical messurements made were fructose, galactose, inositol, total carbohydrate and total protein in seminal plasma and pH in gel-free semen. Fengjing boars had higher concentrations of galactose in seminal plasma than Meishan or Yorkshire boars (P < 0.05), while Meishan seminal plasma had higher galactose concentrations than Yorkshire seminal plasma (P < 0.10). Fructose, inositol and total carbohydrate concentrations were all higher in Fengjing and Meishan seminal plasma than Yorkshire seminal plasma (P < 0.05). Fengjing seminal plasma had higher levels of inositol than Meishan seminal plasma (769 vs 566 mg/100 ml; P < 0.10). Furthermore, Fengjing and Meishan semen had similar fructose and total carbohydrate concentrations (P > 0.80). The pH level and total protein concentration did not differ among the 3 breeds (P > 0.30). This study indicated that breed differences were detected for several seminal parameters, although no single breed had consistantly higher values for all the parameters measured.     

Genetic variability between two breeds based on restriction fragment length polymorphisms (RFLPs) of major histocompatibility complex class I genes in the pig

Summary Restriction fragment length polymorphism analyses of SLA class I genes were performed on 55 Duroc and 24 Hampshire boars from the 1986–87 national performance tests of each breed. Few boars were inbred. Southern blotting and hybridization procedures were performed on genomic DNA isolated from white blood cells by using Pvu II, Bam HI, and Eco RI endonucleases and a swine MHC class I probe. Genetic variability within and between the two breeds was estimated in terms of nucleotide diversity, by using a mathematical analysis based on the different RFLP patterns. The nucleotide diversity calculated within each breed was less than that between the two breeds. The results from the nucleotide diversity analysis suggested that genetic variability was greater in the Duroc breed than in the Hampshire breed. A relatively high level of genetic variability was shown in the class I major histocompatibility complex genes in the pig.     

Within-breed heterozygosity of canine single nucleotide polymorphisms identified by across-breed comparison

Identification of single nucleotide polymorphisms (SNPs) by DNA sequence comparison across breeds is a strategy for developing genetic markers that are useful for many breeds. However, the heterozygosity of SNPs identified in this way might be severely reduced within breeds by inbreeding or genetic drift in the small effective population size of a breed (population subdivision). The effect of inbreeding and population subdivision on heterozygosity of SNPs in dog breeds has never been investigated in a systematic way. We determined the genotypes of dogs from three divergent breeds for SNPs in four canine genes (ACTC, LMNA, SCGB, and TYMS) identified by across-breed DNA sequence comparison, and compared the genotype frequencies to those expected under Hardy-Weinberg equilibrium (HWE). Although population subdivision significantly skewed allele frequencies across breeds for two of the SNPs, the deviations of observed heterozygosities compared with those expected within breeds were minimal. These results indicate that across-breed DNA sequence comparison is a reasonable strategy for identifying SNPs that are useful within many canine breeds.     

Genetic polymorphism in sheep plasma detected using antibodies to human plasminogen

Protein variation was identified in sheep when Western blots of polyacrylamide gels (routinely used to resolve transferrin polymorphism) were stained using antibodies to human plasminogen. The affinity of the antibodies to ovine plasma was less than 7% that of a human standard but they bound specifically to a single polymorphic protein. In 146 lambs and their parents the inheritance of the ovine plasminogen antigen polymorphism was consistent with four autosomal alleles segregating codominantly. However, an additional two lambs had types which were incompatible with their putative parents. The pedigrees of these lambs were tested by DNA fingerprinting and shown to have been incorrectly recorded. The genetic polymorphism detected by human plasminogen antiserum provided a probability of sire exclusion (PE) ranging from 0.04 to 0.32 and a polymorphic information content (PIC) of 0.08 to 0.50 in flocks of five sheep breeds: Perendale, Romney, Merino, Texel and Coopworth (in order of increasing genetic variation in this locus). Significant differences in allele frequency were observed between breeds but sampling did not assess the variation among flocks within a breed.