西藏牛亚科部分群体线粒体DNA遗传多样性研究

为了在从母系遗传上研究西藏牛亚科部分群体的遗传多样性和亲缘进化关系,通过PCR扩增西藏巴美牛、当地土种牛、驼峰牛以及引进娟珊牛4个群体18个个体的mtDNA D-loop控制区全序列,并测序.结果显示,D-loop区大小分布在892 bp～912 bp之间,不同群体间存在长度差异,同时西藏各群体牛D-loop区富含A、T碱基,共检测到颠换、转换、插入、缺失以及颠换和转换共存同一位点等5种突变类型.4个群体发现17种单倍型,多态位点达到125个,平均单倍型多样性约为0.992,平均核苷酸多样性约为0.039,表明西藏黄牛群体遗传多样性丰富.系统进化分析显示17种单倍型大致可以分为三大类,在其进化过程中出现相互交流的情况,巴美牛和驼峰牛亲缘关系最近,当地土种牛仍为较纯的黄牛,西藏黄牛群体不包含瘤牛单倍型,均为普通牛单倍型.     

野牦牛与其他牛科动物GHRHR基因部分序列的比较

对野牦牛生长激素释放激素受体（GHRHR）基因部分片段（包括部分外显子6和部分内含子6）进行PCR扩增、克隆和序列测定,序列已提交到GenBank中（GenBank Accession No：EU872256）。利用BioEdit7.0.9.0、DnaSP 4.10.9等生物信息学软件对野牦牛该基因序列与GenBank中普通牛、瘤牛、水牛、绵羊等牛科动物相应序列进行比对分析。结果表明：野牦牛与普通牛、瘤牛、水牛、绵羊4个物种基因序列间同源性大小依次为99.0%、98.3%、97.8%、92.0%,5个物种间共发现42处序列间碱基差异（9.84/100 bp）,其中野牦牛与普通牛、瘤牛、水牛、绵羊4个牛科物种均存在差异碱基3处;推导的部分氨基酸序列间同源性大小依次为94.4%、94.4%、100.0%、94.4%。     

牛亚科3个主要家养牛种MyoG基因多态性及其遗传分化

对普通牛(鲁西牛、渤海黑牛)、天祝牦牛和中国广西水牛的MyoG基因部分核苷酸序列变异进行了分析,旨在揭示不同牛种群的DNA多态性及其遗传分化.采用PCR和DNA直接测序技术获得4个牛群体的MyoG基因exon 1和5’侧翼部分序列,与GenBanK公布的牛亚科动物同源序列作比对分析.结果显示,在渤海黑牛和牦牛未发现多态位点;鲁西牛存在1个多态位点,定义了2个单倍型;水牛有6个突变位点,但群内只发现1个多态位点.核苷核苷酸多样性(Pi)在0.000 00～0.001 35之间,说明群体内遗传多态性程度较低。核苷酸歧义度(Dxy)以水牛与鲁西牛之间比较最高,为0.013 97,表明水牛遗传分化明显.种系进化树分析表明,牦牛与普通牛亲缘关系很近,二者与水牛关系较远,支持将牦牛归为牛属(Bos)的一个亚属或一个种的观点.     

基于mtDNAcyt b基因变异的中国黄牛品种间遗传关系分析

18个品种136个个体的mtDNA cyt b基因全序列不存在长度差异(1140 bp),均以ATG为起始密码子,中间无插入、缺失或终止密码子,以终止密码子AGA结束;一共检测到了105个替代突变位点,约占核苷酸总数(1140 bp)的9.2％.核苷酸多样性(Pi)为0.00923,一共定义了47个cytb基因单倍型,单倍型多样性为0.848.105个突变位点中有65个转换和39个颠换,参考序列V00654的15615位点为中国黄牛cytb基因变异的转换与颠换共存位点.北方黄牛(0.00550±0.00088)和南方品种(0.00655±0.00128)的平均遗传距离相对较小,但是两者之间的平均遗传距离(0.01271±0.00262)差异较大;中原牛种群和北方种群之间的遗传分化程度(0.01002±0.00193)要大于和南方种群之间的遗传分化程度(0.00891±0.00178),所以中原牛种群受南方瘤牛的遗传影响较大.中原牛种群内的晋南牛和秦川牛受北方普通的影响相对瘤牛要大;而鲁西牛、郏县红牛和南阳牛受南方瘤牛的影响较大.     

牦牛GPRIN3基因的克隆及组织表达分析

为探索GPRIN3基因的生物学特性,以麦洼牦牛为试验材料,利用RT-PCR技术扩增GPRIN3基因编码区序列并对其进行生物信息学分析,采用实时荧光定量PCR技术检测GPRIN3在牦牛不同组织的mRNA表达水平.结果表明,克隆得到GPRIN3基因编码区序列大小为2 343 bp,编码780个氨基酸残基;麦洼牦牛与野牦牛氨基酸序列相比,一致性为99.62%,共有3个位点发生突变,分别是103(G→R)、366(T→M)、404(E→K).系统进化分析显示,牦牛与野牦牛亲缘关系最近,其次是普通牛,与黑猩猩的亲缘关系最远;含有GRIN-C超家族保守功能结构域;实时荧光定量PCR结果表明,GPRIN3在麦洼牦牛肺脏组织中表达量最高,其次是肝脏,在臀肌中表达量最低.本研究为进一步探讨牦牛GPRIN3基因及其编码蛋白的结构和功能提供理论基础,为牦牛分子育种提供新思路.     

四川西部黄牛的染色体核型与分类初探

采用外周血淋巴细胞培养,气干法制备染色体标本,对川西11个县的地方黄牛抽样进行染色体核型及带型研究并与黑白花奶牛、西门塔尔牛、秦川牛等普通牛(Bos tautus)的核型比较,结果表明:川西地方黄牛在遗传基础上和瘤牛(Bos indicus)基本一致而和普通牛存在显著差异:川西地方黄牛y染色体为近端着丝粒染色体(Acroccntric)普通牛y染色体为亚中着丝粒染色体(Submetacentric);核糖体基因的位置和数目三种牛之间也不相同;外部形态观察结果川西牛具有瘤牛的三种瘤峰、较发达的垂皮和体格特征,在生物学特性方面具有瘤牛的耐热、耐粗饲、抗湿、抗蜱等特点,根据上述细胞遗传学证据、结合形态学、生物学特性的研究认为:川西地方黄牛属Bos属瘤牛种(Bos indicus),对川西瘤牛的发生及演化作了初步探讨。     

青海省曲麻莱牦牛的母系遗传多样性及遗传背景探究

为从分子水平上揭示青海省曲麻莱牦牛的母系遗传多样性及遗传背景,对34头曲麻莱牦牛线粒体D-loop区的部分序列进行测定,确定多态位点、单倍型数目、核苷酸多样度及单倍型多样度,并进行系统发育分析。结果表明:在获得的618 bp D-loop序列中,排除1处插入/缺失后,共检测到36处多态位点,其中单一多态位点4处,简约多态位点32处;根据序列间的核苷酸变异共确定了20种单倍型,其中优势单倍型为H1,单倍型多样度(h)为0.952±0.021,核苷酸多样度(π)为0.018±0.009,将测得结果与中国牦牛进行对比,曲麻莱牦牛具有非常丰富的母系遗传多样性。使用NJ法构建的系统发育树结果显示,20种单倍型可以分为2个支系,表明曲麻莱牦牛有2个母系起源。     

牦牛kappa-酪蛋白基因第4外显子的克隆测序及多态性研究

依据普通牛的kappa-酪蛋白基因序列设计引物进行PCR扩增,测定了牦牛kappa-酪蛋白基因第4外显子全序列及部分第4内含子序列,用HindⅢ和PstⅠ切割PCR产物发现,在牦牛群体中具有和普通牛相同的单倍体型.进一步对60个牦牛个体进行群体分析后发现,牦牛群体中的基因频率和基因型频率与普通牛的群体有着很大的差异.     

牛年说奇牛

2009年是我国农历的已丑年,俗称牛年。牛年说牛,别有趣味。《简明不列颠百科全书》记载:牛是驯化的牛科大角型哺乳动物,曾经成群地活动在北美、欧洲、亚洲和非洲。现代的牛分为黄牛、瘤牛、水牛、牦牛、奶牛、杂种牛等多种。然而,在牛的世界里,还存在一些奇牛和怪牛。     

从LDH同工酶的分析探讨野牦牛的育种价值

采用聚丙酰胺凝胶电泳法分离野牦牛血清LDH同工酶并对之作理化分析,发现它具有H型亚基占相对优势的酶谱分布特征,各同工酶在单位时间内的活力、作用pH区限、热稳定性及对尿素变性剂的抗力显著不同,测定4种牛血清LDH的活力结果表明,野牦牛的总活力和比活力最大,次为犏牛,黄牛,而牦牛最低,结果为犏牛杂优效应分析和牦牛育种方案——导入野牦牛血液(基因)提高家牦牛生产性能的研究提供了有意义的生化学基础。     

九龙牦牛肉质性状测定及肌肉蛋白质双向电泳的初步分析

肉质性状是畜禽最主要的经济性状.为阐明九龙牦牛(Bos grunniens)肉质的特性,对不同年龄和性别的九龙牦牛(n=107)肉的部分性状进行了测定.结果表明,0.5岁犊牦牛背最长肌的蛋白质、肌内脂肪(IMF)和肌红蛋白(Mb)含量、剪切力等均显著低于成年牦牛,而这些指标在成年公牦牛与成年母牦牛之间差异不大.不同年龄成年九龙牦牛背最长肌IMF含量接近,提示其IMF沉积受年龄影响相对较小.另外,牦牛背最长肌剪切力存在较大的个体间差异.与成年黄牛相比,牦牛背最长肌IMP含量低,蛋白质含量高,熟化后p H下降幅度小.本研究还建立了九龙牦牛肌肉双向电泳方法,为阐明肌肉差异表达蛋白提供了方法.九龙牦牛肉的蛋白质、IMF和Mb含量以及剪切力等与黄牛有明显差异.     

Mitochondrial DNA analysis of Bronze Age horses recovered from Chifeng region, Inner Mongolia, China

In this study, mitochondrial DNA (mtDNA) analysis was carried out on 9 Bronze Age horses recovered from Dashanqian and Jinggouzi archaeological sites in Chifeng region, Inner Mongolia. China to explore the origin of Chinese domestic horses. Both mtDNA 16S rRNA gene and control region (D-loop) fragments of ancient horses were amplified and sequenced. The analysis of the highly conservative 16S rRNA gene sequences indicated that the burial environment of Chifeng region is suitable for the preservation of ancient DNA (aDNA). Combing 465 mtDNA D-loop sequences representing different breeds from East Asia, Central Asia, Near East and Europe, we constructed a phylogenetic network to investigate the relationship between ancient and modern horses. The phylogenetic network showed that the 9 horses were distributed into different modern horse clusters which were closely related to them representing a certain geographical distribution. Our results showed that the maternal genetic line of the ancient horses in Chifeng region was highly diversified, which contributed to the gene pool of modern domestic horses and suggested a complex origin of domestic horses in China.     

Mitochondrial DNA analysis of Bronze Age horses recovered from Chifeng region, Inner Mongolia, China

In this study, mitochondrial DNA (mtDNA) analysis was carried out on 9 Bronze Age horses recovered from Dashanqian and Jinggouzi archaeological sites in Chifeng region, Inner Mongolia. China to explore the origin of Chinese domestic horses. Both mtDNA 16S rRNA gene and control region (D-loop) fragments of ancient horses were amplified and sequenced. The analysis of the highly conservative 16S rRNA gene sequences indicated that the burial environment of Chifeng region is suitable for the preservation of ancient DNA (aDNA). Combing 465 mtDNA D-loop sequences representing different breeds from East Asia, Central Asia, Near East and Europe, we constructed a phylogenetic network to investigate the relationship between ancient and modern horses. The phylogenetic network showed that the 9 horses were distributed into different modern horse clusters which were closely related to them representing a certain geographical distribution. Our results showed that the maternal genetic line of the ancient horses in Chifeng region was highly diversified, which contributed to the gene pool of modern domestic horses and suggested a complex origin of domestic horses in China.     

Genetic polymorphisms of milk epithelial mucin of yaks

Milk epithelial mucin (MUC1) of yaks was separated by SDS-polyacrylamide gel electrophoresis. A total of nine types of MUC1 were revealed in 427 yaks from five yak breeds including Maiwa yak, Jiulong yak, Tianzhu white yak, Qinghai yak and Tibetan yak. The molecular weights of MUC1 are from 163 kD to 208 kD, most of which are larger than those of bovine. Population genetic analysis shows that the gene frequency and genotype frequency of yak MUC1 differ among breeds, with relatively high gene heterozygosity. The five yak breeds studied fall into two groups according to their milk MUC1 gene frequency, suggesting that milk MUC1 exhibits specificities for local yak groups.     

Genetic evidence for Near-Eastern origins of European cattle

The limited ranges of the wild progenitors of many of the primary European domestic species point to their origins further east in Anatolia or the fertile crescent. The wild ox (Bos primigenius), however, ranged widely and it is unknown whether it was domesticated within Europe as one feature of a local contribution to the farming economy. Here we examine mitochondrial DNA control-region sequence variation from 392 extant animals sampled from Europe, Africa and the Near East, and compare this with data from four extinct British wild oxen. The ancient sequences cluster tightly in a phylogenetic analysis and are clearly distinct from modern cattle. Network analysis of modern Bos taurus identifies four star-like clusters of haplotypes, with intra-cluster diversities that approximate to that expected from the time depth of domestic history. Notably, one of these clusters predominates in Europe and is one of three encountered at substantial frequency in the Near East. In contrast, African diversity is almost exclusively composed of a separate haplogroup, which is encountered only rarely elsewhere. These data provide strong support for a derived Near-Eastern origin for European cattle.     

薄层色谱扫描法测定鹿龟酒中栀子甙的含量

用薄层扫描法测定鹿龟酒中栀子甙的含量，加样回收率为９８．５４％，ＲＳＤ＝２．３％（ｎ＝６）．     

西藏黄牛血清淀粉酶（Am）、酯酶（Es）同工酶遗传特性的研究

应用聚丙烯酰胺凝胶电泳法研究了西藏黄牛的Ａｍ和Ｅｓ同工酶，得出了其基因频率和基因型频率．结果表明，西藏黄牛的Ａｍ和Ｅｓ两个同工酶位点的基因分布及其频率与瘤牛种（Ｂｏｓｉｎｄｉｃｕｓ）差异较大，而与普通牛种（Ｂｏｓｔａｕｒｕｓ）较为一致，这进一步从同工酶位点上验证了我们对西藏黄牛的Ｙ染色体特征和血液蛋白多态性的研究结果     

牦牛血清蛋白质的标定及应用

采用3种测定方法分析野牦牛、家牦牛和犏牛血清蛋白质的各组分含量、分子量及水解氨基酸、游离氨基酸的含量,比较其异同点,结果发现:野、家牦牛具有基本相似的血清蛋白质组成,而γ-球蛋白占总球蛋白的比值、个别蛋白带分子量和氨基酸组成含量等方面有某些显著的差异,表明青海牦牛血清蛋白质发生了不同于野牦牛的遗传变异,在牦牛品种培育中利用两个参数(γ-球蛋白占总球蛋白的比值和血清中必需氨基酸与非必需氨基酸之比值)作为选种标志有定向育种效果。