微卫星标记OarHH35,OarHH55和BM1329与乐至黑山羊产羔数相关性研究

根据微卫星标记在相近物种中具有高度保守性的特点,采用比较基因组学方法,初步探讨了3个与绵羊、山羊产羔率紧密相关的微卫星标记OarHH35、OarHH55和BM1329在4代85只乐至黑山羊中与其产羔数的相关性.结果显示微卫星标记BM1329在乐至黑山羊中可检测到的等位基因数为3,其片段大小分别为125、147和300 bp;标记OarHH55可检测到的等位基因数为2,片段大小分别为106和181 bp;标记OarHH35可检测到的等位基因数为3,片段大小分别为88、112和175 bp.统计分析表明,BM1329的300 bp等位基因和OarHH35的112 bp等位基因对乐至黑山羊产羔数呈正效应(P<0.05),而BM1329的147 bp等位基因和OarHH35的175 bp等位基因对产羔数呈负效应(P<0.05).因此,微卫星标记OarHH35和BM1329可作为乐至黑山羊高繁殖性能候选标记进行深入研究.     

微卫星标记OarAE101和BM1329承五个绵羊品种中的初步研究

选择与Booroola羊高繁殖力主效基因FecB紧密连锁的两个微卫星标记OarAE101和BM1329,分析其在小尾寒羊、湖羊、夏洛来羊、乌珠穆沁羊、多赛特羊和多赛特公羊×小尾寒羊母羊杂一代中的多态分布情况.结果表明微卫星标记OarAE101在5个绵羊品种中的等位基因数为5或4,BM1329在5个绵羊品种中的等位基因数均为4,OarAE101/BM1329在小尾寒羊、湖羊、夏洛来羊、乌珠穆沁羊、多赛特羊和多赛特公羊×小尾寒羊母羊杂一代中的多态信息含量值分别为0.57/0.54、0.62/0.67、0.61/0.59、0.62/0.66、0.56/0.67和0.62/0.68.小尾寒羊OarAE101基因型为107bp/113bp所对应的产羔数最小二乘平均值显著高于基因型为109bp/109bp和107bp/111bp所对应的最小二乘平均值(P＜0.05),产羔数最小二乘平均值在OarAE101其余基因型之间没有显著差异;OarAE101等位基因107bp与小尾寒羊产羔数有显著正相关,109bp和111bp与小尾寒羊产羔数有显著负相关.小尾寒羊BM1329基因型为146bp/158bp所对应的产羔数最小二乘平均值都显著高于其余3种基因型所对应的最小二乘平均值(P＜0.05),产羔数最小二乘平均值在BM1329其余3种基因型之间没有显著差异;BM1329等位基因146bp与小尾寒羊产羔数有显著正相关,148bp与小尾寒羊产羔数有显著负相关.     

小尾寒羊五个微卫星基因座遗传多态性研究

小尾寒羊是我国优良的地方绵羊品种,具有极高的繁殖力,平均每胎产羔2．6只。利用与绵羊高繁殖力主效基因Fec^B和FecX^1连锁的5个微卫星标记（OarAE101,BM1329,BMS2508,TGLA54t TGLA68)对244小尾寒羊母羊进行了遗传检测。用非变性（中性）聚丙烯酰胺凝胶电泳检测同卫星的PCR扩增产物,计算了5个同卫星基因座的等位基因频率,多态信息含量,基因纯合度和杂合度。在小尾寒羊中检测到BM1329有6个等位基因,片段大小为160－180bp,164bp等位基因频率最高（0．6320）;检测到OarAE101有9个等位基因,片段大小为97－135bp,97bp等位基因频率最高（0．7930）;检测到TGLA54有5个等位基因,片段大小为116－136bp,134bp等位基因频率最高（0．8500）;检测到TGLA68有2个等位基因,片段大小为98－100bp,2个等位基因频率相近,检测到BMS2508有6个等位基因,片段大小为93－115bp,99bp等位基因频率最高（0．4795）。BM1329,OarAE101,TGLA54,TGLA68,BMS2508的多态信息含量／基因纯合度／杂合度分别为0．4481／0．4840．0．5160,0．3516／0．6375／0．3625,0．2528／0．7326／0．2674,0．3733／0．5034／0．4966,0．5809／0．3581／0．6419。可见BMS2508的遗传变异最大,TGLA54的遗传变异最小。这些结果可为小尾寒羊种质特性研究提供分子基础数据。     

可可西里自然保护区藏羚羊的微卫星多态性研究

藏羚羊是我国特有的珍稀濒危动物,对其开展遗传多样性的研究具有非常重要的科学价值。为了获取足够的遗传信息并进一步研究藏羚羊在核基因水平上的遗传多样性,对来自可可西里地区的75个藏羚羊干皮张样本进行了微卫星遗传多样性研究。研究从来自牛和绵羊的25个微卫星基因座中筛选到9个具有高度多态性的微卫星基因座(MCM38,MNS64,IOBT395,MCMAI,TGLA68,BM1329,BMS1341,BM3501和MB066)。用非变性聚丙烯凝胶电泳检测微卫星的PCR扩增产物,计算了这9个微卫星基因座的等位基因频率、多态信息含量、基因杂合度等指标并估算了种群数量。结果在75只藏羚羊中共检测到85个等位基因,9个微卫星基因座的等位基因数为7~12个,平均每个基因座检测到9.4个等位基因,有效等位基因数为处于4.676~9.169之间,平均为6.519;基因频率分布在0.007~0.313之间,多态信息含量在0.753~0.881之间,平均为0.818;观察杂合度为0.791~0.897,平均为0.844,期望杂合度为0.786~0.891之间,平均为0.838±0.0132,各基因座观察杂合度与期望杂合度比较接近。固定指数为-0.269~-0.097,平均为-0.163。Shannon’s指数为1.660~2.315,平均为1.990。种群数量的估算结果显示这75个体均来自同一种群。结果表明该种群在核基因水平仍具有丰富的遗传多样性。     

绵羊18号染色体微卫星多态性与后臀发育关系的研究

根据绵羊18号染色体遗传连锁图谱及相关报道, 选择了可能与Callipyge基因连锁和有关的10个高度多态微卫星基因座(ILSTS54、TGLA337、 HH47、TGLA122、BP33、OB2、BM3413、MCM38、MCMA26和CSSM18), 分别对Dorset和Suffolk两个肉羊群体的遗传特性进行了分析。研究发现, 同一微卫星基因座在Suffolk与Dorset群体间的群体遗传参数以及对同一性状的影响情况明显不同。Dorset群体中, 等位基因个数为8~16个, 杂合度为0.8370~0.9252, 多态信息含量为0.8221~0.9167; 而在Suffolk群体中相应参数分别为5~10, 0.7603~0.8913与0.7176~0.8809。对10个微卫星基因座进行臀宽的最小二乘分析, 结果表明: 在Dorset群体中, 微卫星基因座BM3413、MCMA26和CSSM18对臀宽有显著影响(P<0.05), 其余7个微卫星对后臀宽度的影响均未达到显著水平(P>0.05); Suffolk群体中微卫星基因座TGLA122、BM3413、MCM38和CSSM18对绵羊臀宽存在显著或极显著影响(P<0.05或0.01), 其余6个微卫星基因座对臀宽的影响均未达到显著水平(P>0.05)。研究表明, 导致新疆肉羊群体后臀肌过度发育可能是其他基因或QTL的作用。     

湖羊6号染色体微卫星标记多样性与产羔数的关系

选择位于绵羊6号染色体上FecB基因紧密连锁的6个微卫星标记, 即LSCV043、BMS2508、GC101、300U、Bulge5和471U, 分析其在湖羊中的遗传多样性以及和产羔数的关系。结果表明, 6个微卫星位点均属多态性位点, 共检测到34个等位基因、53种基因型。LSCV043、BMS2508和300U属高度多态位点, 其多态信息含量分别为0.6674、0.6035和0.5615。对不同基因型群体的产羔率进行统计分析, LSCV043基因型为107 bp/123 bp所对应的总体产羔数明显高于110 bp/123 bp基因型群体所对应的产羔数(P<0.05); BMS2508基因型为154 bp/154 bp、154 bp/170 bp和154 bp/200 bp所对应的群体第一胎产羔数明显高于170 bp/170 bp所对应的产羔数(P<0.05), 基因型170 bp/170 bp的群体总体产羔数均低于该位点的其他基因型(P<0.05); 其他位点各基因型之间产羔数均无显著差异。     

奶牛微卫星基因座与产奶性能关系的研究

根据小鼠与牛的遗传比较图谱及相关报道选择了与weaver基因连锁的7个微卫星基因库,并在荷斯坦牛群体中对这些基因座进行群体遗传学特性分析。选择具有中度以上多态性的4个微卫星基因座（BM6438、BMS2321、BMS711和TGLA116）进行产奶性能的相关分析。最小二乘分析结果表明,4个微卫星基因座对产奶量影响不显著（P＞0．05）,BM6438生BMS711基因座对乳成分影响不显著（P＞0．05）,BMS2321基因座对乳蛋白量和乳蛋白率有极显著的影响（P＜0．01）,TGLA116对乳蛋白量和乳蛋白率的影响达到0．05的显著水平。     

秦川母牛群体遗传特性的微卫星标记研究

为了从DNA分子水平揭示秦川牛群体遗传多态性和群体遗传结构,寻找可用于秦川牛的微卫星标记,本研究选择了12个普通牛（Bos taurus）微卫星标记检测了90头秦川母牛各微卫星位点的遗传变异及多态性。结果表明,在秦川母牛群体中,12个微卫星位点共检测到了247个等位基因,各位点的等位基因数在13（INRA005）～33个（HEL13）之间,平均每个微卫星位点的等位基因数为21个;总有效等位基因数和平均每个位点平均有效等位基因数（Ne）分别分为142．6229和11．8852。各位点平均基因频率取样方差（V（pij））为2．6036×10^-4。12个微卫星位点平均观察杂合度（Ho）和平均期望杂合度（He）在0．7842（INRA005）～0．9775（BM315）和0．7952（BM315）～0．9446（HEL13）之间。12个位点平均多态信息含量（PIC）在0．7653（INRA005）～0．9420（HEL13）之间,平均为0．8965．12个微卫星位点均属于高度多态位点,这表明秦川母牛群体中所检测各微卫星位点具有丰富的遗传多态性,具备较大的选择潜力。12个微卫星位点的平均固定指数（F）为-0．0076,即各位点杂合子的缺陷度不高,即偏离Hardy—Weinberg平衡的程度不大。     

中国美利奴(新疆军垦型)绵羊9个微卫星基因座多态性研究

利用PCR技术和复合电泳银染技术检测中国美利奴(新疆军垦型)绵羊第1号染色体上BM6506,BM1824,BM6438, ILSTS004和OarDB6 等5个基因座和第6号染色体上 BM4621,OarHH55,BM143和OarJMP8 等4个基因座,共9个基因座的基因频率（Pi）、个体鉴别力（DP）、杂合度（H）、多态信息含量（PIC）、和非父排除概率（PE）。结果显示：9个微卫星基因座的基因型分布符合Hardy-Weinberg平衡,绵羊中9个微卫星基因座中BM4621 基因座的DP、H、PIC和PE都为最高。9个微卫星基因座的累积个体鉴别力(CDP)为0.99999,累积非父排除能力(CPE)为0.99915。结果显示9个微卫星基因座适用于中国美利奴(新疆军垦型)绵羊的遗传连锁分析、个体识别和亲权鉴定等研究领域。     

小尾寒羊微卫星与RAPD标记的研究

小尾寒羊是世界上具有非季节性发情和多胎特性的高繁殖率绵羊品种之一。选择4个位于绵羊6号染色体上且与FecB基因紧密连锁的微卫星标记,对小尾寒羊基因组进行PCR扩增后,采用最小二乘法估计各等位基因片段对产羔数的影响。分析结果表明,等位基因OarJIA-5、OarJIA-10、BM143-12和OarHH55-11可以作为小尾寒羊多胎位点的分子标记。从100条随机引物中筛选出18条引物,对小尾寒羊、大尾寒羊、洼地绵羊、滩羊等4个绵羊品种和鲁北山羊的基因组进行扩增,共扩增出146条带,其中94条表现出多态性,占64．60%,同时扩增出每个品种的特异性条带。采用Nei氏标准距离和NJ聚类分析对不同品种的遗传关系进行分析,结果表明,4个绵羊品种亲缘关系很近,提示它们可能起源于共同的原始祖先。     

Genetic variability among Polish Red, Hereford and Holstein-Friesian cattle raised in Poland based on analysis of microsatellite DNA sequences

Polymorphism of 11 microsatellite DNA loci was analysed in Polish Red (PR), Hereford and Holstein-Friesian (HF) cattle raised in Poland and genetic distance among these breeds was determined. At the 11 loci (TGLA227, BM2113, TGLA53, ETH10, SPS115, TGLA126, TGLA122, INRA23, ETH3, ETH225 and BM1824) analysed with automated DNA sizing technology, a total of 213 alleles were identified: 76 in PR, 76 in HF, and 61 in Hereford. All the microsatellite DNA markers showed high polymorphism. Polymorphism information content (PIC) calculated for each marker exceeded 0.5, except for the ETH3 locus in Hereford cattle (PIC=0.475), and heterozygosity (H) ranged from 54.1% to as much as 85.2%. The coefficient of genetic distance was 0.354 between PR and Hereford, 0.414 between HF and Hereford, and 0.416 between PR and HF cattle.     

A proposal for standardization in forensic bovine DNA typing: allele nomenclature of 16 cattle-specific short tandem repeat loci

In this study, a proposal is presented for the allele nomenclature of 16 polymorphic short tandem repeat (STR) loci ( BM1824 , BM2113 , ETH10 , ETH225 , INRA023 , SPS115 , TGLA122 , TGLA126 , TGLA227 , ETH3 , TGLA53 , BM1818 , CSRM60 , CSSM66 , HAUT27 and ILSTS006 ) for bovine genotyping ( Bos taurus ). The nomenclature is based on sequence data of the polymorphic region(s) of the STR loci as recommended by the DNA commission of the International Society of Forensic Genetics for human DNA typing. To cover commonly and rarely occurring alleles, a selection of animals homozygous for the alleles at these STR loci were analysed and subjected to sequence studies. The alleles of the STR loci consisted either of simple or compound dinucleotide repeat patterns. Only a limited number of alleles with the same fragment size showed different repeat structures. The allele designation described here was based on the number of repeats including all variable regions within the amplified fragment. The set of 16 STR markers should be propagated for the use in all bovine applications including forensic analysis.     

Individual identification of Chinese Holstein Bull by 10 STR loci

With the premium bull plays a growing important role in cattle industry, semen detection technology based on individual identification and phylogenetic relationship is paid more and more attention. In order to lay the foundation for the establishment of the China Holstein bull identification method, this research takes 20 Chinese Holstein dairy bull’s blood and their corresponding semen, and then extracts the DNA both from the blood and semen, analysis the genetic polymorphisms of 10 microsatellite loci (TGLA227, INRA23, TGLA122, BM2113, SPS115, ETH3, ETH225, MCM158, MAF45 and UMN0108) by microsatellite marker, discuss the feasibility of this method used to individual identification. The results showed that Chinese Holstein dairy bull genetic diversity in the ten microsatellite loci were both high, and the average polymorphic information content of TGLA227, which highest, is 0.8162, ETH225 has the lowest, which is 0.6224. Use STR loci to identify the bull’s semen, the cumulative individual identification capacity is 99.99 %, which indication that 10 STR loci can be used to the frozen semen quality test and cows individual identification.     

Determination of ancestral proportions in synthetic bovine breeds using commonly employed microsatellite markers

The International Society of Animal Genetics (ISAG) has chosen nine microsatellites (international marker set) as a standard that should be included in all cattle parentage studies. They are BM1824, BM2113, INRA023, SPS115, TGLA122, TGLA126, TGLA227, ETH10, and ETH225. We decided to ascertain whether this microsatellite set could be used to determine ancestral proportions in individual animals of synthetic breeds produced by crossing zebu and taurine cattle. Since the genotypes of these markers are routinely available, this would constitute a practical and cost-free method to estimate the ancestry of synthetic breed animals. Genotypes of 100 Gir and 100 Holstein animals were examined for this ISAG marker set. As expected, there were very significant allele frequency differences between the two breeds at most loci. We also typed 20 Girolando animals for which there was complete genealogical information. "Structure" software easily distinguished Holstein and Gir animals based on their microsatellite genotypes; it also attributed the genomic proportion of zebu and taurine of each of the 20 Girolando animals. The proportion of Holstein ancestry was then regressed on the genealogical data; there was a highly significant correlation (r = 0.84, P < 0.0001). The nine microsatellites that compose the ISAG international marker set were capable of estimating the ancestral Gir and Holstein genomic proportions in individual Girolando animals within narrow confidence limits. This microsatellite set might also be useful for estimating the proportions of taurine and zebu origins in commercial meat products.     

Application of bovine microsatellite markers for genetic diversity analysis of Swiss yak (Poephagus grunniens)

In order to assess the applicability of bovine microsatellite markers for population genetic studies in Swiss yak, 131 bovine microsatellite markers were tested on a panel of 10 animals. Efficient amplification was observed for 124 markers (94.6%) with a total of 476 alleles, of which 117 markers (94.3%) were polymorphic. The number of alleles per locus among the polymorphic markers ranged from two to nine. Seven loci (ILSTS005, BMS424B, BMS1825, BMS672, BM1314, ETH123 and BM6017) failed to amplify yak genomic DNA. Two cattle Y-chromosome specific microsatellite markers (INRA126 and BM861) amplified genomic DNA from both male and female yaks. However, two additional markers on cattle Y-chromosome (INRA124 and INRA189) amplified DNA from only males. Of the polymorphic markers, 24 microsatellites proposed by CaDBase for within- and cross-species comparisons and two additional highly polymorphic markers (MHCII and TGLA73) were used to investigate the genetic variability and the population structure of a Swiss yak herd that included 51 additional animals. The polymorphic information content ranged from 0.355 to 0.752, while observed heterozygosity (HO) ranged from 0.348 to 0.823. Furthermore, a set of 13 markers, organized into three multiplex polymerase chain reactions, was evaluated for routine parentage testing. This set provided an exclusion probability in a family of four yaks (both parents and two offspring) of 0.995. These microsatellites serve as useful tools for genetic characterization of the yak, which continues to be an important domestic livestock species.     

Identification of fecundity gene (FecB) carriers using microsatellite markers and its effect on sheep weight

Abstract. Altogether 115 animals representing 5 genetic groups: 3 purebred Booroola Merino, Corriedale and Olkuska, and 2 Booroola crossbreds were included in the studies. In total 6 alleles from 97 bp to 119 bp in microsatellite OarAE101, and 5 alleles from 162 bp to 174 bp in BM1329 were identified. The marker of FecB gene presence seems to be an allele of 97 bp in the case of microsatellite OarAE 101 and 162 bp in the case of BM1329. Significant differences FecB carriers (Booroola-Corriedale) and non-carriers (Corriedale) in birth weight and at weaning at 100 days (males and females from twins) as well as weight gain during the first 28 days and 100 days were found. Purebred lambs showed higher values of the investigated traits.     

Genetic variation at microsatellite loci in Spanish sheep

Genetic variation at 18 microsatellite loci was analysed in six indigenous Spanish sheep: Churra; Latxa; Manchega; Rasa-Aragonesa; Castellana and Merino. Merinos had frequently the highest number of alleles per locus, whereas Latxas showed the lowest one at many loci. Markers ordered decreasingly according to the number of variants differentiated in the whole population were: MAF70; TGLA13; CSSM66; BM143, BM6444; MAF36; MAF64; CSSM6; TGLA53; OarFCB11; MAF33; BM4621; MAF48; MAF65; BM1258; ILSTS002; ADCYC and OarCP34. Parameters of variability such as effective number of alleles and gene diversities corroborated the high level of variation frequently displayed by microsatellite markers. Comparison of allele distributions among populations and loci did not reveal consistent shapes. Distributions were centralised in some cases, whereas in others some kind of skewness was evident. Breed-specific alleles were detected at most loci, being frequent in Merinos and rare in Churras.