贵州四个山羊品种mtDNA多态性及起源分化

采用１５种限制性内切酶,研究贵州省４个山羊品种共９３只个体的线粒体ＤＮＡ多态性。其中ＢｏｍＨＩ、ＨｉｎｄⅢ和ＳａｌⅠ３种酶的酶切类型存在多态。共检测到１８种限制性态型,归结为３种ｍｔＤＮＡ单倍型。单倍型Ⅰ、Ⅱ在贵州山羊４个品种分布频率较高,分别为７７．４２％和２１．５０％,单倍型Ⅲ分布频率较低（１．０８％）;品种间亲缘关系聚类分析表明白山羊和黑山羊亲缘关系最近,其次为黔林羊,而与小香羊的亲缘关系最     

青海湖裸鲤mtDNA遗传多样性的初步研究

本文用BclⅠ、AvaⅠ、BamHⅠ、PstⅠ、KpnⅠ、PvuⅡ共6种限制性内切核酸酶,分析了15尾青海湖裸鲤mtDNA的限制性片段长度多态性,共检测出20个酶切位点,发现BclⅠ、BamHⅠ和PvuⅡ三种酶切类型具有多态性.根据不同个体mtDNA的酶切类型,青海湖裸鲤存在4种mtDNA单倍型,计算mtDNA多态度π值为0.0043,初步认为青海湖裸鲤在线粒体DNA上存在较丰富的群体内变异、 Abstract:An analysis of patterns of cleavage of mtDNA by restriction endonucleases was performed for 15 Gymnocypris przewalskii przewalskii.Mitochondrial DNA polymorphisms were detected in the restriction patterns generated by the following 5 enzymes,BclI,AvaI,BamH],PstI,KpnI,PvuII.Only 3 of them (BclI,BantHI,PvuII) were found to be polymorphisms.Our results shorted that there were 4 mtDNA haplotypes in Gyrnnocypris przewalskii przewalskii and the genetic divergenec(π)was 0.0043,which indicated that mtDNA genetic diversity in Gymnocypris przewalskii przewalskii is higher.     

青海湖裸鲤mtDNA遗传多样性的初步研究

本用Bcl、AvaⅠ,BamHⅠ,PstⅠ,KpnⅠ,PvuⅡ共6种限制性内切核酶,分析了15尾青海湖裸鲤mtDNA的限制片段长度多态性,共检测出20个酶切位点,发现BclⅠ,BamHⅠ和PvuⅡ三种酶切类型具有多态性。根据不同个体mtDNA的酶切类,青海湖裸鲤存在4种mtDNA单位型,计算mtDNA多态度π值为0．0043,初步认为青海湖裸鲤在线粒体DNA上存在较丰富的群体内变异。     

山羊品种间线粒体DNA限制性片段长度多态性研究

本试验利用ＡｐａⅠ、ＢａｍＨⅠ、ＢｃｌⅠ、ＢｇｌⅠ、ＢｇｌⅡ、ＣｌａⅠ、ＥｃｏＲⅠ、ＥｃｏＲⅤ、ＨａｅⅡ、ＨｉｎｄⅢ、ＫｐｎⅠＰｓｔⅠ、ＰｖｕⅡ、ＳａｃⅠ、ＳａｌⅠ、ＳｍａⅠ和ＸｈｏⅠ计１８种限制性内切酶,研究了来自欧洲,非洲及国内的５个山羊品种共计３３只个体的ｍｔＤＮＡ,共检测了２７处限制性态型,可归结为８种单倍型。结果表明,国内２个百品种的基本单倍型为ＢａｍＨⅠ－Ａ,ＢｃｌⅠ－Ｂ、ＣｌａⅠ－Ａ     

大尺蠖核多角体病毒DNA的限制性消化和物理图谱

用5种限制性核酸内切酶消化大尺蠖核多角体病毒(BsNPV)基因组DNA,所得片段数分别是:BamH Ⅰ,9;Bgl Ⅱ,7;Xho Ⅰ,10;HindⅢ 13;EcoR Ⅰ,12,从各个片段的累加测出BsNPV基因组的平均大小约为91,75kb;分子量约为59.60×10~6d。在单酶消化的基础上,用BamH Ⅰ/Bgl Ⅱ双酶消化得到16个片段(即9+7);大小为91,27kb,用、Bgl Ⅱ/Xho Ⅰ双酶消化产生7+10=17个片段,大小为90.04kb,由此组建了这三个酶在BsNPV基因组上的物理图谱。     

民猪的SM-DRB基因的PCR-RFLP多态性分析

采用克隆测序和PCR—RFLP相结合的方法,对民猪的SLA—DRB基因的整个编码区进行了扫描和多态性分析．结果表明该基因的第2外显子是整个基因的高变区,突变率达到5．6％,其中80％为有效突变,但没有发现新的等位基因;PCR-RFLP结果表明外显子1用内切酶Alu Ⅰ酶切可获得3种基因型;外显子2用内切酶TaqⅠ酶切可获得3种基因型：外显子3用内切酶BcnⅠ酶切可获得3种基因型：外显子4用内切酶MboⅠ酶切可获得2种基因型：外显子5用内切酶HinlⅠ酶切可获得3种基因型．Hardy-Weinberg平衡分析表明民猪在Alu Ⅰ和HinlⅠ处于平衡状态（P〉0．05）．在TaqⅠ、BcnⅠ和MboⅠ处于不平衡状态．     

细胞质雄性不育水稻线粒体基因组的RFLP分析

利用RFLP技术,比较研究了在农业生产上广泛应用的9种细胞质雄性不育体系的线粒体基因组,结果表明：1）9种水稻雄性不育细胞质的遗传相似性变化范围为0．615～1．000。所有配子体雄性不育细胞质的遗传相似性变化范围为0．6431．000,其中QXA,DY1A和YM15A这3种细胞质的遗传相似性为1．000。所有孢子体雄性不育细胞质的遗传相似性为1．000,2）在配子体细胞质雄性不育水稻中,用3个探针（atpa,atp9＋nad6,cox2）与2种内切酶（HindⅢ,BamHⅠ）的组合未发现不育系与保持系之间的多态性,但在探针atp6,cob,和had2的杂交带型中找到了不育系和保持系之间的一些差异。YTA和YTB在5个探针．内切酶组合（atp61HindⅢ,coblHindⅢ,atp61BamHⅠ,coblBamHⅠ,nad21BamHⅠ）中存在差异。3种细胞质（QX,SJ,DY1）的不育系和保持系之间的差异是相同的,都出现在atp6／HindⅢ,atp6／BamHⅠ,cob／BamHⅠ等3个组合中。YM15A和YM15B在4个组合（atp6／HindⅢ,atp6／BamHⅠ,cob／BamHⅠ,nad2／BamHⅠ）中存在差异。LYA和LYB的差异出现在cob／HindⅢ,cob／BamHⅠ,nad2／BamHⅠ这3个组合中;3）在孢子体细胞质雄性不育水稻中,所有不育系的带型是一样的,所有保持系的带型也一样。不育系和保持系的差异出现在atp6／HindⅢ,cob／HindⅢ,atp6／EcoR,Ⅰcob／EcoRⅠ,cox1／EcoRⅠ,atp6／BamHⅠ,cob／BamHⅠ,cox1／BamHⅠ,cox2／BamHⅠ等9个组合中。这些结果在分子水平上揭示了9种雄性不育细胞质的线粒体基因组存在结构多样性,并为其细胞质雄性不育分子机理的研究打下了基础。     

民猪的SLA-DRB基因的PCR-RFLP多态性分析

采用克隆测序和PCR-RFLP相结合的方法,对民猪的SLA-DRB基因的整个编码区进行了扫描和多态性分析.结果表明该基因的第2外显子是整个基因的高变区,突变率达到5.6%,其中80%为有效突变,但没有发现新的等位基因;PCR-RFLP结果表明外显子1用内切酶Alu Ⅰ酶切可获得3种基因型;外显子2用内切酶Taq Ⅰ酶切可获得3种基因型;外显子3用内切酶Bcn Ⅰ酶切可获得3种基因型;外显子4用内切酶Mbo Ⅰ酶切可获得2种基因型;外显子5用内切酶Hin1 Ⅰ酶切可获得3种基因型.Hardy-Weinberg平衡分析表明民猪在Alu Ⅰ和Hin1 Ⅰ处于平衡状态(P＞0.05),在Taq Ⅰ、Bcn Ⅰ和Mbo Ⅰ处于不平衡状态.     

鳜及大眼鳜线粒体DNA比较研究

采用10种限制性内切酶HindⅢ、EcoRⅠ、BglⅠ、BglⅡ、BamHⅠ、PvuⅡ、PstⅠ、SalⅠ、XbaⅠ、XhoⅠ对鳜及大眼鳜肝脏线粒体DNA（mtDNA）进行了分析。鳜鱼mtDNA分子量为9.703×106u,大小为16．19kb;大眼鳜mtDNA分子量为9．603×106u,大小为16．02kb。根据单、双酶切结果构建了鳜及大眼鳜mtDNA10种酶限制性酶切图谱,并对两种鱼的mtDNA酶切图谱进行了比较。     

褐家鼠线粒体DNA遗传多态性的研究

通过碱变性法提取线粒体ＤＮＡ,用地高辛标记的探针Ｓｏｕｔｈｅｒｎ杂交限制性酶切多态性（ＲＦＬＰ）分析,研究中国家鼠Ｒａｔｔｕｓｎｏｒｖｅｇｉｃｕｓ遗传多态性。采用ＡｐａⅠ、ＡｖａⅠ、ＢａｎＨＩ、ＢｃｌⅠ、ＢｇｌⅠ、ＣｌａⅠ、ＥｃｏＲⅠ、ＥｃｏＲⅤ、ｈｉｎｄⅢ、ＰｖａⅡ、ＳｃａⅠ和ＸｂａⅠ等１２种限制性内切酶分析来自我国８个地区２６只褐家鼠的线粒体ＤＮＡ,共检出２０种限制性态型和１１种ｍｔＤＮＡ单倍     

10个牛品种线粒体12S rRNA基因多态性分析

以鲁西牛、渤海黑牛、大别山牛、南阳牛、晋南牛、秦川牛、中国西门塔尔牛、日本和牛、海福特牛和安格斯牛10个牛品种共计353头牛为研究对象,利用单链构象多态性（SSCP）分子标记方法对其线粒体DNA 12S rRNA基因进行了多态性分析,结果发现该基因出现分布频率不同的3种单倍型（Ⅰ、Ⅱ、Ⅲ）,其中单倍型Ⅰ出现的频率最低,仅在晋南牛1个个体中发现;单倍型Ⅱ在中国本地黄牛中都存在,在中国西门塔尔牛中的分布频率很低,但在3个国外品种海福特牛、安格斯牛和日本和牛中不存在;单倍型Ⅲ在这10个牛品种中均有分布,但以国外品种最丰富。计算平均多态信息含量,发现我国地方品种在0.232～0.423之间,基本属于中度多态,说明我国地方黄牛品种在12S rRNA上多态性比较丰富,国外品种较为贫乏;但是单倍型Ⅰ频率很低有丢失的趋势,需要对其进行保护。     

Mitochondrial DNA variation in cattle of South China: origin and introgression

Ten restriction endonucleases were used to investigate the mitochondrial DNA restriction fragment length polymorphism (mtDNA RFLP) of 11 native cattle breeds and one cultivated cattle breed in South China. Twenty-three restriction morphs were detected, which can be sorted into five haplotypes. A phylogenetic tree of the haplotypes was constructed by using the 'upgMa' method. Our study showed that haplotype I and II are identical to the zebu (Bos indicus) and taurine (Bos taurus) haplotypes, respectively. Zebu and taurine were the two major origins of cattle populations in South China, and the zebu probably had more influence on the native cattle population than taurine did. Haplotype III is identical to haplotype I of yak (Bos grunniens), which was only detected in the Diqing cattle breed. Haplotype IV was detected for the first time. This haplotype, found only in Dehong cattle, might be from an independent domestication event, probably from another Bos indicus population. Divergence of haplotypes I and IV occurred about 268,000-535,000 years ago, much earlier than the 10,000-year history of cattle husbandry. Our results also suggest a secondary introgession of mtDNA from yak to Diqing cattle.     

Analysis of MHC class I genes across horse MHC haplotypes

The genomic sequences of 15 horse major histocompatibility complex (MHC) class I genes and a collection of MHC class I homozygous horses of five different haplotypes were used to investigate the genomic structure and polymorphism of the equine MHC. A combination of conserved and locus-specific primers was used to amplify horse MHC class I genes with classical and nonclassical characteristics. Multiple clones from each haplotype identified three to five classical sequences per homozygous animal and two to three nonclassical sequences. Phylogenetic analysis was applied to these sequences, and groups were identified which appear to be allelic series, but some sequences were left ungrouped. Sequences determined from MHC class I heterozygous horses and previously described MHC class I sequences were then added, representing a total of ten horse MHC haplotypes. These results were consistent with those obtained from the MHC homozygous horses alone, and 30 classical sequences were assigned to four previously confirmed loci and three new provisional loci. The nonclassical genes had few alleles and the classical genes had higher levels of allelic polymorphism. Alleles for two classical loci with the expected pattern of polymorphism were found in the majority of haplotypes tested, but alleles at two other commonly detected loci had more variation outside of the hypervariable region than within. Our data indicate that the equine major histocompatibility complex is characterized by variation in the complement of class I genes expressed in different haplotypes in addition to the expected allelic polymorphism within loci.     

Chinese Mongolian horses may retain early domestic male genetic lineages yet to be discovered

The Mongolian horse represents one of the most ancient extant horse populations. In this study we determined the male‐specific region of the Y chromosome (MSY) haplotype distribution in 60 Chinese Mongolian horses representing five distinct populations. Cosmopolitan male lineages were predominant in horses from one improved (Sanhe), one Chinese Mongolian subtype (Baicha Iron Hoof) and one indigenous (Abaga Black) population. In contrast, autochthonous Y chromosome diversity was evident among the two landrace populations (Wushen and Wuzhumuqin), as the majority of their MSY haplotypes were situated at root nodes in a network. Our results also suggest gene flow between Chinese Mongolian and Arabian horses, as an appreciable number of Wuzhumuqin horses carried haplotypes that are typically observed in Arabian horses. Although most horses carried modern haplotypes as a direct result of recent breed improvement, authentic Chinese Mongolian horses retain an ancient signature of paternal lineages that has not previously been described in extant horse populations. Therefore, further characterization of MSY variation in these populations will be important for the discovery of lost diversity in modern domestic horses and also for understanding the evolutionary history of equine paternal lineages.     

Mitochondrial DNA polymorphism of a triploid form of Fasciola in Japan

Mitochondrial DNA polymorphism was characterized in a triploid form of Fasciola found in Japan in comparison with F. hepatica, F. gigantica and Korean Fasciola worm. Seventy worms of Fasciola from Japan, three of F. hepatica from Uruguay and Australia, two of F. gigantica from Thailand and one of Fasciola from Korea were used in the study. Mitochondrial DNA polymorphism was detected by restriction fragment length polymorphism (RFLP) using eight restriction enzymes, BamH I, Bgl II, Dra I, EcoR I, EcoR V, Hind III, Mfl I and Sca I. Three different types (types 1, 2 and 3) were detected from 76 Fasciola worms used in the study. Eight of 70 Japanese worms were categorized in type 2 (F. gigantica type), and the remaining 62 were in type 3 (F. hepatica type).     

我国主要地方绵羊品种mtDNA D-loop区PCR-RFLP研究

利用5种限制性内切酶（Hinf I,Msp I,Sau3A I,Xsp I,Taq I）,采用PCR-RFLP技术研究了我国9个地方绵羊品种以及2个引入品种共计83只绵羊个体线粒体DNA D-loop区的多态性。结果表明,我国主要地方绵羊品种线粒体DNA D-loop区存在两种基本单体型,提示我国主要地方绵羊品种起源于两个母系祖先。线粒体DNA D-loop区多态度为0.042 1%,说明我国地方绵羊品种线粒体DNA多态度较为贫乏。
     

Association of genetic polymorphisms in vitamin D receptor gene and susceptibility to sporadic prostate cancer

Genetic and environmental factors are involved in prostate cancer (PCa) etiology. Single nucleotide polymorphisms (SNPs) may contribute to the PCa pathogenesis. The goal of this study is to determine the role of vitamin D receptor (VDR) gene polymorphisms and haplotypes in the development and progression of sporadic PCa. One hundred and thirty-three PCa patients and 157 age-matched healthy controls were genotyped for the Apa I (rs7975232), Bsm I (rs1544410) and Taq I (rs731236) polymorphisms in VDR gene by using polymerase chain reaction-restriction fragment length polymorphism. An association was observed between the Apa I polymorphism and PCa predisposition (P = 0.03). When compared with AA genotype, there was a highly notable difference in the frequencies of the Aa (P = 0.02), aa (P = 0.026) and Apa I 'a' allele carriers (Aa + aa) (P = 0.009) genotypes. Furthermore, we found a statistical difference in the allele frequencies of the Apa I polymorphism between the sporadic PCa patients and control subjects (P = 0.013). The genotype distribution for the Bsm I and Taq I polymorphisms were similar between cases and controls (P > 0.05). No clinically significant relationship was found between the three-locus haplotypes and development of sporadic PCa. The genotype frequencies for the three polymorphisms of the VDR gene within subgroups of PCa (defined by tumor stage, Gleason score, PSA levels) were also analyzed, but no statistically noteworthy difference was observed (P > 0.05). As far as we know, this is the first study which investigates the relationship between VDR genotypes and sporadic PCa in the Turkish population. Our findings suggest that the VDR ApaI (rs7975232) polymorphism may play a role in the development of sporadic PCa.     

Maternal and paternal genetic variation in Estonian local horse breeds in the context of geographically adjacent and distant Eurasian breeds

The maternal and paternal genetic variation of horse breeds from the Baltic Sea region, including three local Estonian breeds, was assessed and compared with that of Altai and Yakutian horses. In the mtDNA D‐loop region, 72 haplotypes assigned to 20 haplogroups in the nine breeds were detected. In Estonian local breeds, 38 mtDNA haplotypes were found, and five of them were shared by the three breeds. More than 60% of all identified haplotypes were rare. Compared with the Estonian Native and Estonian Heavy Draught breeds, a higher haplotypic diversity was found in the Tori breed (h = 0.969). Moreover, four haplotypes shared among Finnish and Estonian local horse breeds indicated ancient ancestry, and of these, H30 (haplogroup D3) showed global sharing and genetic links between modern Baltic Sea region and Siberian horses, specifically. The studied breed set showed high variability in maternal inheritance and mixed patterns of the international and native breeds of the Siberian and Baltic regions. No variation was found in paternally inherited markers among horse breeds in the Baltic Sea region.     

Haplotype diversity of the myostatin gene among beef cattle breeds

A total of 678 individuals from 28 European bovine breeds were both phenotyped and analysed at the myostatin locus by the Single Strand Conformation Polymorphism (SSCP) method. Seven new mutations were identified which contribute to the high polymorphism (1 SNP every 100 bp) present in this small gene; twenty haplotypes were described and a genotyping method was set up using the Oligonucleotide Ligation Assay (OLA) method. Some haplotypes appeared to be exclusive to a particular breed; this was the case for 5 in the Charolaise (involving mutation Q204X) and 7 in the Maine-Anjou (involving mutation E226X). The relationships between the different haplotypes were studied, thus allowing to test the earlier hypothesis on the origin of muscular hypertrophy in Europe: muscular hypertrophy (namely nt821(del11)) was mainly spread in different waves from northern Europe milk purpose populations in most breeds; however, other mutations (mostly disruptive) arose in a single breed, were highly selected and have since scarcely evolved to other populations.