蒙古绵羊和哈萨克绵羊MHC-DRB3基因外显子2的多态性

采用PCR RFLP方法对蒙古绵羊和哈萨克绵羊MHC DRB3 基因第 2外显子 2 85bp的扩增产物进行多态性分析 ,共检测到 17种基因型 ,由A、B、C、D、E、F和H共 7个复等位基因控制。通过酶切图谱分析表明 ,蒙古绵羊和哈萨克绵羊的MHC DRB3 基因第 2外显子的第 15 4、16 8和 2 2 0位的碱基表现出多态性。统计分析表明 ,MHC DRB3 基因的部分基因型频率和等位基因频率在两个群体之间差异显著或极显著 (P <0 10、P <0 0 5或P <0 0 1)。χ2 适合性检验结果表明 ,蒙古绵羊和哈萨克绵羊的MHC DRB3 基因第 2外显子的HaeⅢ酶切位点均未达到Hardy Weinberg平衡状态 (P <0 0 1)。     

蒙古山羊和哈萨克山羊GOLA-DRB3基因的HaeⅢ酶切多态性分析

采用限制性内切核酸酶HaeⅢ对蒙古山羊和哈萨克山羊GOLA-DRB3基因外显子2的285bp扩增产物进行了PCR-RFLP多态性分析,共检测到17种基因型,由A、B、C、D、E、F和H等7个复等位基因控制;通过酶切图谱分析发现蒙古山羊和哈萨克山羊的GOLA-DRB3基因外显子2的154、168和220位碱基表现出多态性。并对基因型频率和等位基因频率进行了统计分析,结果表明,GOLA-DRB3基因的部分基因型频率和等位基因频率在两个群体之间差异显著（P＜0.10或P＜0.05）或极显著（P＜0.01）; χ2适合性检验结果表明,蒙古山羊和哈萨克山羊的GOLA-DRB3基因外显子2的HaeⅢ酶切位点均未达到Hardy-Weinberg平衡状态（P＜0.01）。 Abstract:The exon2 of GOLA-DRB3 gene was amplified and a uniform fragment of 285bp was obtained in Mongolian Goat and Kazakh Goat.The 285bp PCR product was digested with restriction endomuclease HaeⅢ and genetic polymorphism was investigated by PCR-RFLP.Seventeen kinds of genotypes were found in two populations,which were controlled by seven alleles.There are significant differences in some genotypic frequencies and gene frequencies between the two populations（P＜0.10,P＜0.05,P＜0.01）;The results of χ2 test showed that genotypes of GOLA-DRB3 gene in two populations did not fit with Hardy-Weinberg equilibrium（P＜0.01）.     

猪MyoG基因的PCR-RFLP多态性分析

以杜洛克、长白、大约克、南昌白、二花脸、梅山猪、玉山黑猪、乐平花猪、金华两头乌及上高两头乌等中外10个猪种共计561头猪为研究材料,采用3对引物（PCR1、PCR2、PCR3）分别扩增猪肌细胞生成素（MyoG）基因的不同区域,扩增产物经限制性核酸内切酶MspⅠ酶切后发现：（1）在PCR1 MspⅠ-RFLP位点上,外来品种杜洛克、长白、大约克及培育品种南昌白中极大多数个体表现为AA型,个别为BB型;而6个中国地方猪种除乐平花猪外均以BB型居多。（2）在PCR2 MspⅠ-RFLP位点上,6个中国地方猪种除一头玉山黑猪表现为MN型外,其余均为MM型;而外来品种以NN型占大多数,培育品种南昌白更趋向于外来品种。（3）在PCR3 MspⅠ-RFLP位点上,所有猪种均可得到扩增产物,但无MspⅠ酶切位点。（4）在梅山猪及与其亲缘关系较近的二花脸猪中,没有发现Soumillion等(1997)报道的梅山猪特异性MspⅠ多态性酶切位点。     

用4个微卫星标记分析7个绵羊群体之间的遗传关系

分析了4个微卫星基因座BM143、OarHH35、OarAE101、BMS2508在7个绵羊群体（小尾寒羊、湖羊、乌珠穆沁羊、萨福克羊、多赛特羊、夏洛来羊、多赛特公羊×小尾寒羊母羊F1代杂种羊）286只绵羊中的遗传多态性。结果表明,这4个微卫星标记在7个绵羊群体中的等位基因数分别为9、11、14和9,其多态信息含量/有效等位基因数/杂合度分别为0.7073/3.7231/0.7314、0.8267/6.4399/0.8447、0.5743/2.5178/0.6028、0.6172/3.0712/0.6744,其中OarHH35的遗传变异最大,OarAE101最小。7个绵羊群体中小尾寒羊的遗传变异最大,湖羊的最小。基于Nei氏DA距离和DS标准遗传距离,采用UPGMA方法构建了系统发生树。该发生树将中国地方品种（小尾寒羊、乌珠穆沁羊、湖羊）和法国的夏洛来羊归为一类,将F1杂种羊、英国品种（萨福克羊和多赛特羊）归为另一类。绵羊微卫星基因分型技术为检查品种（群体）之间的遗传关系提供了一个有用的工具。 Abstract：The genetic polymorphisms of four microsatellite loci BM143,OarHH35,OarAE101,and BMS2508 were analyzed in 286 sheep of seven sheep populations (Small Tail Han sheep, Hu sheep, Ujumqin sheep, Suffolk sheep, Dorset sheep, Charolais sheep, F1 of Dorset♂ × Small Tail Han sheep♀). The numbers of alleles for BM143,OarHH35,OarAE101,and BMS2508 are 9, 11, 14 and 9 in seven sheep populations, respectively. The polymorphism information content/number of effective alleles/ heterozygosity of BM143,OarHH35,OarAE101 and BMS2508 were 0.7073/3.7231/0.7314, 0.8267/6.4399/0.8447,0.5743/2.5178/0.6028,0.6172/3.0712/0.6744 in 286 sheep, respectively. The results revealed the greatest genetic variation at OarHH35 locus and the lowest at OarAE101, the greatest genetic variation in Small Tail Han sheep and the lowest in Hu sheep among seven sheep populations. In the unweighted pair group method with arithmetic mean (UPGMA) dendrograms based on Nei's DA distance and Nei's DS standard genetic distance, the Chinese native breeds (Small Tail Han sheep, Ujumqin sheep, Hu sheep) were grouped together, then with Charolais sheep. The F1 crossbred sheep, and the two British native sheep (Suffolk sheep, Dorset sheep) also clustered together. Microsatellite genotyping in sheep provided a useful tool for examining the genetic relationships among breeds(populations).     

多浪羊mtDNA D-loop区遗传多样性及系统发育分析

以多浪羊为研究对象,分析绵羊线粒体D-loop区的遗传多样性,为研究多浪羊的起源和进化历史奠定基础。结果显示,多浪羊线粒体DNA D-loop序列长度为945~1 039 bp,A、T、G和C含量分别为29.4%、27.7%、17.7%和25.1%,其中A+T为57.1%,G+C为42.8%。研究获得了26种单倍型,56个多态位点,其中单一多态位点42个,简约信息位点14个。平均核苷酸差异数k为5.289,核苷酸多样度Pi为0.02 415,核苷酸多样度较低,说明多浪羊的遗传多样性贫乏,应采取重点措施予以保护。另外研究发现多浪羊经历过群体扩张,其母系起源除A、B和C世系外,可能存在D或E世系。     

麦洼牦牛乳酸脱氢酶-B基因多态性的PCR-RFLP分析

为了从分子水平上检测麦洼牦牛Bos grunniens乳酸脱氢酶-1(LDH1)的H亚基编码基因Ldhb的多态性,实验提取79头麦洼牦牛基因组DNA,采用PCR-RFLP技术分析Ldhb基因多态性,并测定牦牛背最长肌中肌红蛋白含量。实验建立的牦牛Ldhb的G896A突变位点的PCR-RFLP检测方法,在79头麦洼牦牛中检测到Ldhb-AA和Ldhb-AG两种基因型,其中AG基因型频率为16.46%,并且这些样品的乳酸脱氢酶-1的电泳迁移率快于Ldhb-AA样品。本实验未检测到Ldhb-GG基因型;麦洼牦牛Ldhb基因多态性与背最长肌中的肌红蛋白含量之间未见相关性。     

卡拉库尔羊MHC-DRB3基因的遗传多样性研究

目的:实验通过MCH-DRB3基因来探讨卡拉库尔羊的遗传多态性,为绵羊遗传资源的合理利用及保护提供理论基础和科学依据.方法:采用PCR-Clone测序方法首次对卡拉库尔羊的MHC-DRB3基因的第二外显子进行分子遗传多态性检测与分析.结果:总共检测出35种单倍型,总的单倍型多样度(Hd)为0.958,核苷酸多样性(Pi)为0.05786,平均核苷酸差异数(k)为16.49062,简约信息多态位点数为54.卡拉库尔羊MHC-DRB3基因的氨基酸序列的氨基酸组成进行分析可知含量最多的氨基酸是精氨酸(Arg),平均含量为12.9%,含量最低的氨基酸是蛋氨酸(Met),平均含量为0.018%.结论:从以上参数可以看出卡拉库尔羊具有很高的遗传多样性.     

西藏小型猪SLA-DQA基因外显子2的PCR-RFLP多态性分析

目的分析西藏小型猪SLA-DQA基因第2外显子不同酶切位点的基因型多态性以及等位基因的多态性,检验这些酶切位点上的基因频率是否达到Hardy-Weiberg平衡态。方法采用EcoRⅠ和AluⅠ两种限制性内切酶对西藏小型猪SLA-DQA目的基因的第1和第2内含子部分序列以及完整的外显子2进行PCR-RFLP分析。结果经EcoRⅠ酶切后,以纯合子BB基因型居多,BB、AB、AA基因型频率分布为45.000%、31.667%和23.333%,其中B为优势等位基因（60.833%）;经AluⅠ酶切后,MN基因型频率（50.000%）分别高于MM型（30.000%）和NN型（20.000%）,     

北京地区汉族人群21号染色体上5个STR基因座的遗传多态性

阐明21号染色体上唐氏综合征关键区域内或附近的5个STR基因座（D21S1413、D21S1446、D21S1437、D21S1411、D21S1412）在北京地区汉族人群中的结构特征和群体遗传学数据。Chelex法提取血DNA,PCR扩增后,应用聚丙烯酰胺凝胶电泳和银染法或基因片段扫描检测法进行STR分型,测序后确定STR基因座的主型和进行等位基因的命名。结果该5个STR基因座具有简单重复序列和遗传多态性,杂合度和多态信息含量高。它为唐氏综合征的基因诊断和产前基因诊断提供理论依据,也为这些遗传标记在我国人群中进行亲子鉴定和个体识别提供概率计算依据。Abstract: To elucidate the genetic polymorphisms of five STR loci on chromosome 21 in Chinese Han population and construct a preliminary database,EDTA-blood specimens were collected from unrelated individuals in Beijing. The DNAs were extracted with Chelex method and were amplified by PCR. The PCR products were analyzed by the PAG electrophoresis or by the approach of the automated fluorescent detection. The five STR loci consist of simple repeat motif and its distributions of genotypes are agreement with Hardy–Weinberg equation. Its polymorphism information content is all over 0.50. The obtained data can not only be used as evidences for genetic diagnosis of Down Syndrome, but also for calculating the probabilities in the paternity test and individual identification.     

北京地区汉族群体D1S1612和D18S535基因座的遗传多态性

采用扩增片段长度多态性（Amp-FLP）分型技术,调查中国北京地区汉族群体D1S1612、D18S535 基因座的遗传多态性,获得等位基因频率分布。结果显示, D1S1612检出9个等位基因,25种基因型, D18S535检出9个等位基因,27种基因型。两个STR基因座的杂和度（H）分别为0.779、0.887;个人识别率（Dp）分别为0.901、0.927;非父排除率（PE）分别为0.564、0.770;多态信息容量（PIC）分别为0.723、0.796,卡方检验表明两个STR 基因座基因型频率分布符合Hardy-Weinberg平衡 (P>0.01 )。D1S1612和D18S535 基因座均属高杂合度、高识别能力的遗传标记,可用于法庭科学亲子鉴定和个人识别。 Abstract: To investigate the genetic polymorphism of D1S1612 and D18S535 in Han population of Beijing. Amp-FLP method was used. 9 alleles, 25 genotypes were observed for D1S1612 locus; and 9 alleles and 27 genotypes for D18S535 locus. All allele frequencies, heterozygosity (H), discrimination power (Dp), exclusion of paternity probability (PE) and polymorphism information content (PIC) were calculated. The allele distributions of the two loci were conformed to Hardy-Weinberg equilibrium (P>0.01). According to the results obtained in this study, it is suggested that both D1S1612 and D18S535 are useful genetic markers for individual identification and paternity testing in forensic science practice as well for genetic study.     

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.     

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．     

西藏小型猪H-FABP基因的PCR-RFLP研究

目的研究西藏小型猪心脏脂肪酸结合蛋白（H-FABP）基因5’-上游区和第二内含子内的遗传变异。方法应用PCR-RFLP技术测定30头西藏小型猪H-FABP的基因型。结果（1）在5’-上游区的Hinf I-RFLP位点上,西藏小型猪表现出多态性,等位基因h的频率为0．80;（2）在在第二内含子内的Hae Ⅲ-RFLP位点上,西藏小型猪均为DD纯合子;（3）在第二内含子内的Hinf I＊-RFLP位点上,除一头猪表现为bb基因型外,其余猪都表现为BB基因型,等位基因B的频率为0．97;（4）除Hae Ⅲ-RFLP位点外,在其余位点上西藏小型猪均处于Hardy-weinberg不平衡状态。（5）在Hinf I-RFLP中,西藏小型猪表现为中度多态性（0．25〈PIC〈0．5）,而在其他位点的RFLP中表现为低度多态（PIC〈0．25）。结论可以利用西藏小型猪H-FABP基因5’-上游区的多态遗传标记来分析其与肌内脂肪的关系。     

山西猪种及其杂种群体H-FABP基因的PCR-RFLP研究

利用PCR－RFLP技术对马身猪、山西白猪及其杂种群体共286头猪的心脏脂肪酸结合蛋白（H-FABP）基因5′－上游区（HinfⅠ-RFLP）和第二内含子内（HinfⅠ*-RFLP和Hae Ⅲ-RFLP）的遗传变异进行了研究。结果表明：（1）在Hae Ⅲ-RFLP位点上,马身猪均为DD纯合子,而其他猪群在此位点上均存在变异,马身猪的杂种群体在该位点上只有两种基因型（DD、Dd）;（2）在5′－上游区的HinfⅠ-RFLP位点上,杜洛克猪×山西白猪的杂种群体只有HH基因型,而其他群体都表现出多态性,马身猪等位基因h的频率为0.9727;（3）在第二内含子内的HinfⅠ*-RFLP位点上,马身猪表现出两种基因型（BB、Bb）,等位基因B的频率为0.9667;（4）在HinfⅠ*-RFLP和Hae Ⅲ-RFLP位点上,所有猪群均处于Hardy –Weinberg 平衡状态。     

Genetic Polymorphism of β-Lactoglobulin Gene in Native Turkish Sheep Breeds

The genetic polymorphism of the β-lactoglobulin gene was investigated in three native Turkish sheep breeds. The study was carried out on 108 sheep (29 Kıvırcık, 38 G?k?eada, and 41 Sakız) by means of PCR-RFLP methods. Two genetic variants (A and B) and three genotypes (AA, AB, and BB) of β-lactoglobulin have been identified. The gene frequencies of β-LG A and B were 0.7759 and 0.2241 in Kıvırcık, 0.7632 and 0.2368 in G?k?eada, and 0.9756 and 0.0244 in Sakız breeds, respectively. The populations were in Hardy–Weinberg equilibrium in all samples from the three breeds.     

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。
     

Extensive polymorphism of the BOLA-DRB3 gene distinguished by PCR-RFLP

A polymerase chain reaction (PCR)-based method is described for typing of alleles of the bovine lymphocyte antigen (BoLA)-DRB3 gene. A total of 30 DRB3 alleles were distinguished by digestion of PCR amplification products of BoLA-DRB3 exon 2 with RsaI, BstYI and HaeIII (PCR-RFLP). All restriction fragment patterns, with the exception of one HaeIII pattern, were consistent with restriction sites that were found among 14 previously sequenced DRB3 alleles. The PCR-RFLP typing method was evaluated on 168 genomic DNA samples collected from animals of 10 cattle breeds, 48 of which were typed in the Fourth International BoLA Workshop for BoLA-DRB and -DQ by conventional restriction fragment length polymorphism (RFLP) analysis using heterologous and homologous DNA probes. Thirty-one DRB/DQ haplotypes containing 23 DRB3 alleles were identified among the 48 workshop animals analysed. Using PCR-RFLP, 11 DRB3 alleles were identified in 18 workshop animals for which DRB RFLPs were not informative. PCR-RFLP typing of additional animals revealed five new DRB3 alleles, of which three contained a putatively located three basepair deletion in the identical position as found for the sequenced allele DRB*2A. PCR-RFLP was shown to be a rapid and sensitive method for the detection of polymorphism in a functionally relevant domain of the BoLA-DRB3 gene and should be useful for studying the evolution of DRB polymorphism in cattle and other Bovidae.