人类3-磷酸甘油脱氢酶1基因第三外显子的单核苷酸多态性研究

通过基因组克隆和测序技术,研究人类3.磷酸甘油脱氢酶基因(glycerol 3-phosphatedehydrogenase 1,简称GPD1)第三外显子的单核苷酸多态性(single nucleotide polymorphism,简称SNP)情况.发现成都地区人类GPD1的第三外显子上存在4个SNP位点,分别为G69A,G81A,A121C和A127T,其中G81A和A121C为已经报道过的SNP,G69A和A127T为新发现的SNP.在这些SNP中,A121C和A127T会引起氨基酸的改变,分别为Thr113Pro和Ile115Leu.通过分子建模的方法分析这两个错义突变对GPD1蛋白分子结构的影响,发现113位氨基酸残基的替换为极性氨基酸苏氨酸和非极性氨基酸脯氨酸之间的转换,而且113位氨基酸残基暴露于蛋白分子表面,所以该突变可能会对蛋白分子的性质和功能造成一定的影响.     

Single nucleotide polymorphism analysis in chicken growth hormone gene and its associations with growth and carcass traits

In this experiment, F2 chicken derived from Broilers crossing to Silky are used to study the effect of growth hormone gene on growth and carcass traits. The partial gene is amplified by two pairs of primers, and single nucleotide polymorphism (SNPs) is detected by the technique of PCR-RFLP (restriction fragment length polymorphism), and then confirmed by DNA sequencing. The mutations are found in intron 3 and intron 4 respectively, and can be clarified by digestion with EcoRVand Msp I. The results of least square analysis indicate that the gene has significant association with some carcass traits, such as breast muscle weight, breast muscle rate, abdominal fat rate, and has no association with other growth and carcass traits, such as live weight, carcass weight, eviscerated yield with giblet, eviscerated yield, leg muscle weight, heart weight, liver weight, abdominal fat weight, chest angle width, head and neck weight,shank and claw weight, wing weight, muscular stomach weight, glandular stomach weight, ovary or testicular weight, shank girth, small intestine length, 1-week body weight, 6-week body weight, 12-week body weight, etc. These results demonstrate that GH gene could be a genetic locus or linked to a major gene significantly affecting the growth and carcass traits in chicken.     

Single nucleotide polymorphism analysis of the encoding region of EX-FABP gene and its association with fattiness trait in chicken

Fattiness is an important parameter to estimate meat quality, which has high heritability. In this experiment, F2 chickens derived from Broilers crossing to Silky were used to study the effect of extracellular fatty acid inding protein (EX-FABP) gene on abdominal fat accumulation. Five exons and partial introns of the gene were amplified by six pairs of primers, and then single nucleotide polymorphisms (SNPs) were detected by single strand conformation polymorphism (SSCP) and subsequently confirmed by sequencing. There were four nucleotides variations found, T-C at 659, C-T at 924, CA-TG at 968 and C-T at 1985 respectively. The result of least square analysis suggests that the chickens with HH genotype defined by the third pair of primers have a lower abdominal fat weight and abdominal fat percentage than the chickens with other genotypes (GG, GI, GH and II). It implies that extracellular fatty acid binding protein gene could be a candidate locus or linked to a major gene that significantly affects abdominal fat traits in chicken.     

Sequences and polymorphisms of exons 3 and 4 in porcine UCP2 gene

Uncoupling proteins are mitochondrial membrane transporters, which regulate metabolic pathways of energy balance, and are associated with biological traits of animal body weight, resting metabolic rates and energy conversion. In this study, a region of the exons 3 and 4 of pig UCP2 gene was cloned and analyzed, and a new single nucleotide polymorphic site was detected by PCR-SSCP in five pig breeds. This newfound polymorphism results from a T to G substitution at the position of nucleotide 272, which is located in intron3.     

三江黄牛Bola-DRB3基因第二外显子的PCR-RFLP多态性研究

用PCR-RFLP方法,对三江黄牛和黑白花奶牛的MHCⅡ类基因座位Bola-DRB3基因第2外显子的遗传样性进行了研究.结果表明:①三江黄牛HaeIII酶切出现3种基因型,即SS、RS、RR,由S和R两个基因控制,RR为优势基因型,R为优势基因;RsaI酶切出现4种基因型,即XX、XM、LM、MM,由X、M和L三个复等位基因控制,LM为优势基因型,M为优势基因;两种内切酶在三江黄牛Bola-DRB3基因的第二个外显子上有150、202、65、180等酶切位点,且4个位点的碱基存在多态性,表现为多碱基突变;经χ2适合性检验,HaeIII和RsaI酶切位点的χ2值分别为7.0802、14.1358,差异显著(p<0.05),说明三江黄牛Bola-DRB3基因第2外显子的4个酶切位点的碱基突变未达到Hardy-Weinberg平衡状态,存在选择、基因突变、近交、遗传漂变等因素的影响.②黑白花奶牛HaeIII酶切出现4种基因型,即JJ、KI、IJ、KJ,由J、K和I三个复等位基因控制,JJ为优势基因型,J为优势基因;RsaI酶切出现3种基因型,即LL、MM、LM,由L和M两个基因控制,LM为优势基因型,M为优势基因;HaeIII和RsaI两种内切酶在黑白花奶牛中也有多个酶切位点,在150、48、65、202位上有碱基多态性,表现为多碱基突变;经χ2适合性检验,RsaI酶切位点的χ2值为3.456,差异不显著(p>0.05),处于Hardy-Weinberg平衡状态;HaeIII酶切位点的χ2值为18.4211,差异显著(p<0.05),未达到Hardy-Weinberg平衡状态.③用两种酶在三江黄牛中共检测到7种基因型,且与黑白花奶牛在酶切位点上具有较多的差异,说明三江黄牛具有丰富的遗传多样性;这一结果与张成忠、赖松家等从血液蛋白、染色体、mtDNA等水平上的研究结果一致,也同三江黄牛的育成历史和所处生态条件一致.综上所述,三江黄牛具有丰富的遗传多样性,今后应加强对三江黄牛遗传多态性以及起源、演化和分类的研究,为这一畜种资源的进一步开发利用和保护提供理论基础.     

ODF1基因多态性与四川地区特发性弱精子症的相关性研究

本研究探讨四川地区外周致密纤维1（outer dense fiber1, ODF1）基因多态性与男性特发性弱精子症的相关性.选取106例特发性弱精子症男性不育患者为病例组和104例精液参数正常的男性为对照组,分析ODF1基因多态性在各组中分布的差异.结果表明：与对照组相比,ODF1基因中27- bp缺失变异体（c.656delACCCCTGCAGCCCCTGCAACCCGTGCA）的频率在特发性弱精子症男性不育组中显著升高（OR=1.783,95%CI=1.178~2.700,P=0.006）,并利用MEGA比对分析后发现有10个氨基酸缺失（C218-P229delinsNPCSPCNPCS）.通过PROVEAN与PMP软件分析,27- bp缺失变异体对蛋白质功能有潜在损害.因此四川地区ODF1基因中27 -bp缺失变异体极有可能与弱精子症男性不育相关.     

Cloning, sequencing and polymorphism analyzing of the exon 4 of the kappa-casein gene in yak

According to the nucleotide sequence of cattle kappa- casein gene, a pair of primers was synthesized to amplify the sequences of the exon 4 and partial intron 4 of this gene in yak. The restriction fragment length polymor¬phisms were identified by the digestions of the PCR product with Hind 111 or Pst I in both populations of cattle and yak, 'but the frequency of the allele and the genotype were significantly different between the two animal populations.     

广东汉族人群TLR9基因单核苷酸多态性及其单倍型(英文)

采用PCR扩增和直接测序法对191例健康且无血缘关系的广东汉族人群筛查了TLR9全基因序列,包括调控区、5′非翻译区、第1,2外显子、内含子及3′非翻译区上所有的单核苷酸多态性(SNP)位点.共检出五个SNP位点,分别为调控区的-1 486 T/C和-1 421 C/T、内含子区的+1174 A/G、第2外显子的+1 387 T/C和+2848 G/A,其中-1421 C/T和+1 387 T/C为首次发现的新位点.连锁不平衡分析表明-486 T/C,1174 A/G以及2848 G/A之间存在紧密连锁,并且涵盖了整个基因区域,形成了一个单倍域.在此基础上运用Hhase软件构建了TLR9基因的单倍型,共得到七种单倍型并模拟了它们可能的分布频率.进一步的中性检验表明TLR9基因在广东汉族人群中符合中性进化模式.     

KIT基因单核苷酸多态性与四川地区男性不育的相关性研究

基于Luminex平台,应用多重等位基因特异性引物延伸反应（allelespecific primer extension, ASPE), 对130例无精症、严重少精症患者与115例男性可育对照者KIT基因的4个单体型标签SNPs(haplotype tagging SNP):rs3819387、rs3796768、rs2237023、rs12646437的基因型进行检测并做关联分析.结果显示病例组与对照组中rs3819387、rs3796768、rs2237023、rs12646437位点基因型频率分     

牛POU1F1-exon2多态性与部分线性和乳用性状相关研究

利用PCR-SSCP技术,研究弗莱维赫牛和蒙贝利亚牛POU1F1-exon2多态性,分析其对部分线性评分性状和乳用性状之间的影响.结果表明,POU1F1-exon2存在两个等位基因——C和T.在该位点上基因频率分别为0.8426/0.1574,0.9032/0.0968.并且这两个群体均处于Hardy-Weinberg平衡状态(P＜0.05).TT基因型体重和体高的均值高于CT和CC基因型均值(P＜0.05),而CT基因型均值高于CC基因型均值(P＜0.05).CC和CT基因型腹围均值显著高于TT基因型均值(P＜0.05).TT基因型强壮度和乳用性均值均显著高于CT基因型均值和CC基因型均值(P＜0.05).TT、CT基因型的产奶量均值显著高于CC基因型均值(P＜0.05).