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1.
野猪、民猪、大白猪μ-钙激活酶基因的变异位点分析   总被引:8,自引:2,他引:6  
杨秀芹  刘惠  郭丽娟  许尧  刘娣 《遗传》2007,29(5):581-586
为了进一步研究CAPN1基因变异与肉嫩度的关系, 寻找与猪嫩度性状相关的分子标记, 对CAPN1基因组进行了克隆、测序, 并利用PCR-SSCP方法对其编码区序列进行了分子扫描, 寻找多态位点, 分析不同基因型在野猪、民猪、大白猪中的种间分布规律。获得了猪CAPN1基因的15个内含子序列; 根据GenBank上提供的CAPN1 CDS及克隆的内含子序列设计了5对多态性引物进行PCR-SSCP分析; 共找到8个SNPs, 其中7个位于外显子上, 1个位于内含子上, 并且外显子上的突变有3个是错义突变, 分别造成了蛋白质多肽链上第54位氨基酸的S/T、第192位氨基酸的G/E、第363位氨基酸的V/I替代; χ2独立性检验表明不同基因型在大白猪与野猪、民猪之间存在着极显著的差异(P<0.01), 野猪和民猪之间除了S1引物3种基因型的分布存在显著差异外(0.010.05)。这些多态位点具有成为分子标记的潜在可能。  相似文献   

2.
采用PCR-SSCP方法检测猪胰岛素样生长因子2(insulin-like growth factor 2,IGF2)基因外显子4b的多态性,并分析其对初生重、断奶重、6月龄重和背膘厚的遗传效应。根据猪IGF2基因的DNA序列(AY044828)设计引物,在exon4b上发现了一个多态性位点,并对纯合子进行测序,发现13位C→A转换,且存在3种基因型(AA、AB、BB)。统计结果表明,3种基因型在各品种中的分布不一致,长白猪与大白猪比较,莱芜猪与大薄莲猪比较,沂蒙黑猪和里岔黑猪比较差异不显著(P>0.05);其他猪种间基因型分布的差异均显著(P<0.05)。固定效应模型分析结果表明,初生重,断奶重和背膘厚基因型间差异显著(P<0.05),而6月龄重基因型间差异不显著(P>0.05)。最小二乘分析结果表明,BB基因型个体同AA和AB、基因型个体比较初生重和断奶重的差异显著(P<0.05),3种基因型在初生重和断奶重的大小排列顺序为AA>AB>BB;AA基因型个体同AB和BB基因型个体比较背膘厚的差异显著(P<0.05),3种基因型在背膘厚的大小排列顺序为BB>AB>AA。因此,推测IGF2基因对个体的生长速度和胴体瘦肉率存在一定的影响,将IGF2基因应用于猪育种过程中的标记辅助选择可以加快猪的育种进程。  相似文献   

3.
采用PCR-SSCP方法对长白猪、大白猪、杜洛克猪、山西黑猪和马身猪共636头猪的肌细胞生成素(Myogenin,简称MyoG)基因3′端的遗传多态性进行检测,分析MyoG基因对猪的初生体质量、断奶体质量、6月龄体质量和背膘厚的影响。根据已发表的猪MyoG基因3′端侧翼序列设计3对引物,发现F1/R1引物对扩增的片段有多态性。统计结果发现:长白、大白、杜洛克猪种B基因为优势基因,其基因频率分别为0.8807、0.7256和0.8581;山西黑猪种A基因为优势基因,其基因频率为0.9359;马身猪种只检测到A基因。χ2独立性检验表明,基因型分布在外来猪种(长白猪、大白猪、杜洛克猪)与地方猪种(山西黑猪、马身猪)间存在极显著差异(P<0.01)。固定效应模型分析结果表明,初生体质量基因型间差异显著(P<0.05),而断奶体质量、6月龄体质量和背膘厚基因型间差异不显著(P>0.05)。最小二乘分析结果表明,BB基因型与其它2种基因型比较有较大的初生质量,同AA和AB型比较差异极显著(P<0.01)。因此,推测MyoG基因对个体的初生体质量存在一定的影响,选择带有B等位基因的个体有望提高个体的初生体质量。  相似文献   

4.
以猪解耦联蛋白基因 3(UCP3)作为控制猪胴体与肉质性状主基因的候选基因。利用直接测序法对 4个品种猪骨骼肌中UCP3基因的部分编码区序列 (第 4外显子部分及第 5、6、7外显子全部片段 )进行比较分析 ,发现3个cSNP位点 ,其中ORF中第 84 2碱基的突变可导致相应编码氨基酸序列的改变 :甲硫氨酸→苏氨酸 ,选取此位点作为猪UCP3基因的多态位点。用PCR SSCP检测方法在 3个品种猪中进行该cSNP位点多态性片段的基因型分型 ,结果显示在 3个猪群中表现出 3种基因型 (AA、AB、BB) ,χ2 独立性检验结果表明 3种基因型在各品种间分布不一致 ,梅山猪同大白、长白猪分别比较差异极显著 (P <0 0 1) ;对大白×梅山资源家系F2 代 139头个体进行了该多态片段的基因型鉴定 ,并对其基因型与所检测个体相应的胴体、肉质性状采用GLM分析进行遗传效应研究 ,结果表明 :该基因对一些胴体、肉质性状有显著性影响 ,并且该基因以加性效应为主 (如 ,眼肌高度、背最长肌色值、系水力的加性效应都达显著水平 )。因此 ,推测UCP3基因可能是影响猪胴体及肉质性状的主效基因或与主效基因紧密连锁的标记基因 ,并且能够在分子标记辅助选择中用于对猪胴体、肉质性状的遗传改良及固定  相似文献   

5.
猪Mx1基因第14外显子多态性分析及新突变位点的 发现   总被引:1,自引:0,他引:1  
采用PCR-RFLP方法对国内外7个猪种Mx1基因第14外显子的多态性进行分析, 共检测到3个等位基因, 6种基因型。其中杜洛克中仅存在AA基因型, 苏太猪中存在全部基因型, 只有在梅山猪和具有梅山猪血统的苏太猪中出现基因型BB。所有猪种中, 只有在地方猪种和培育猪种中出现等位基因B, 所有猪种除松辽黑猪外均以A为优势等位基因。卡方检验结果表明, 不同猪种间基因型分布差异较大, 梅山猪和松辽黑猪与其他所有猪种的基因型频率差异极显著(P<0.01) , 苏太猪与除皮特兰猪外的所有猪种的基因型频率差异也极显著(P<0.01) , 淮猪与杜洛克和约克夏这两个国外猪种基因型频率差异不显著(P>0.05), 而与皮特兰和其他地方猪种的基因型频率均存在极显著差异(P<0.01) 。通过测序在扩增片段中新发现了3种类型的碱基突变, 前2个分别导致了Thr和Glu向Ala和Arg的替换, 最后一个突变不引起氨基酸的变化, 且后两个突变位点为BB基因型所特有。  相似文献   

6.
张增荣  朱庆  蒋小松  杜华锐 《遗传》2007,29(8):982-988
为了探讨CAPN1基因作为影响鸡肌肉嫩度候选基因的可能性, 寻找与鸡嫩度性状相关的分子标记, 对钙蛋白酶Ⅰ(CAPN1)基因的CDS区进行SNPs 检测, 分析不同基因型在5个优质肉鸡纯品系和3个配套系间分布规律。利用测序和单链构象多态(SSCP)的方法进行SNPs 检测和基因型的分析, 计算等位基因频率、各位点多态信息含量。结果发现2546位(位点A) 处发生点突变由C→T和3535位(位点B)处发生点突变由G→A。各位点的3 种基因型与肉鸡生产性状的最小二乘分析结果表明,各位点的各种基因型个体在肌纤维密度和部分屠体性状指标存在显著差异(P< 0.05)。初步推断CAPN1基因可能是影响鸡嫩度性状潜在的主效基因或与主效基因连锁, 并且这些位点具有成为分子标记的潜在可能。  相似文献   

7.
杨秀芹  刘慧  郭丽娟  关庆芝  许尧  刘娣 《遗传》2008,30(6):741-746
以野猪、民猪和大白猪为研究对象, 根据网上公布的序列设计了7对引物, 采用测序、PCR-SSCP和PCR-RFLP方法对CAPN1基因的部分外显子和3′UTR区进行了单核苷酸多态性检测和基因型分析, 探讨CAPN1基因多态性与瘦肉率和嫩度的关系。研究发现11个SNPs, 其中5个位于外显子, 4个位于内含子, 2个位于3′UTR区, 外显子中的突变有一处是错义突变, 导致了蛋白质多肽链第260位氨基酸发生了M/V的替代。群体遗传学分析表明, 在所检测的各多态位点上, 野猪、民猪、大白猪3个品种间不同基因型的分布都存在着极显著的差异(P<0.01), 而野猪和民猪之间各基因型的分布差异不显著(P>0.05), 民猪和大白猪之间各基因型的分布存在着极显著的差异(P<0.01)。结合品种特性分析表明, P4、P6引物和3′ UTR区HinfⅠ位点所检测的不同基因型和瘦肉率具有一定的相关性。  相似文献   

8.
猪IGF2基因的遗传多态性及其遗传效应分析   总被引:9,自引:0,他引:9       下载免费PDF全文
薛慧良  徐来祥 《遗传》2008,30(2):179-179―184
采用PCR-SSCP方法检测胰岛素样生长因子2 (insulin-like growth factor 2, IGF2)基因外显子7, 8, 9的多态性, 并分析其对初生重、断奶重、6月龄重和6月龄背膘厚的遗传效应。根据猪IGF2基因的DNA序列(AY044828)设计3对引物, 结果在Ex8引物对扩增的片段上发现了多态性, 并对纯合子进行测序, 发现exon8的53位存在C→T转换, 且检测到3种基因型(AA、AB、BB)。统计结果表明, 3种基因型在各品种中的分布不一致, 长白猪与大白猪比较, 莱芜猪与大薄莲猪比较, 沂蒙黑猪和里岔黑猪比较差异不显著(P > 0.05); 其他猪种间基因型分布的差异均显著(P < 0.01)。固定效应模型分析结果表明, 初生重和6月龄背膘厚基因型间差异显著(P < 0.05), 而断奶重和6月龄重基因型间差异不显著(P > 0.05)。最小二乘分析结果表明, BB基因型个体同AA和AB基因型个体比较初生重的差异显著(P < 0.05), 3种基因型在初生重的大小排列顺序为AB > AA > BB; AA基因型个体同AB和BB基因型个体比较6月龄背膘厚的差异显著(P < 0.05), 3种基因型在6月龄背膘厚的大小排列顺序为BB > AB > AA。因此, 推测IGF2基因对个体的初生重和胴体瘦肉率存在一定的影响, 将IGF2基因应用于猪育种过程中的标记辅助选择可以加快猪的育种进程。  相似文献   

9.
以野猪.民猪和大白猪为研究对象,根据网上公布的序列设计了7对引物,采用测序,PCR-SSCP和PCR-RFLP方法对CAPN1基因的部分外显子和3'UTR区进行了单核苷酸多态性检测和基因型分析,探讨CAPN1基因多态性与瘦肉率和嫩度的关系.研究发现11个SNPs,其中5个位于外显子,4个位于内含子,2个位于3'UTR区,外显子中的突变有一处是错义突变,导致了蛋白质多肽链第260位氨基酸发生了M/V的替代.群体遗传学分析表明,在所检测的各多态位点上,野猪、民猪、大白猪3个品种间不同基因型的分布都存在着极显著的差异(P<0.01),而野猪和民猪之间各基因型的分布差异不显著(P>0.05),民猪和大白猪之间各基因型的分布存在着极显著的差异(P<0.01).结合品种特性分析表明,P4、P6引物和3'UTR区Hinf1位点所检测的不同基因型和瘦肉率具有一定的相关性.  相似文献   

10.
利用Oligo功能分类基因芯片检测了瘦肉型的长白猪和脂肪型的太湖猪在1、2、3、4和5月龄间背部皮下脂肪中脂肪沉积代谢和细胞生长调控相关基因的动态表达变化。差异表达分析结果显示1~5月龄的品种间分别有10、6、11、8和19个基因的表达差异倍数大于2倍, 且长白猪有25个基因在不同月龄间的表达差异达显著水平(P<0.05)。其中血管生成素样蛋白4 (ANGPTL4)、组织蛋白酶K (CTSK) 、异柠檬酸脱氢酶2(NADP+) (IDH2)、脂蛋白脂酶 (LPL)、苹果酸酶1 (ME1)、 硬酯酰辅酶A去饱和酶 (SCD)和解藕联蛋白2 (UCP2)这7个基因不仅在同月龄的品种间和品种内的不同月龄间差异表达, 主成分分析结果也显示其表达模式明显偏离其他基因, 提示受到了特殊的调控。聚类分析结果显示1~5月龄间长白猪中正调控脂肪酸代谢基因的表达量逐渐上调, 太湖猪中参与细胞生长调控基因的表达量平缓波动且变化幅度相对较小。另外, 5个差异表达基因的荧光定量RT-PCR验证结果均与芯片结果呈正相关趋势。结果成功筛选出了对猪胴体和肉质性状可能具有重要影响并值得深入研究的一些候选基因, 初步揭示了相关基因的表达变化规律, 为了解生长发育过程中脂肪酸合成与水解的动态平衡过程提供了基础数据。  相似文献   

11.
12.
Adenosine monophosphate deaminase (AMPD; EC 3.5.4.6) catalyses the hydrolysis of adenosine monophosphate (AMP) to commensurate amounts of inosine monophosphate (IMP) and ammonia. The production of AMP deaminase in Candida albicans was measured in Lee's medium grown cultures. The highest AMPD activity was observed at 24 h of growth. The enzyme had an optimum pH and temperature at 6-7 and 28 degrees C, respectively. This enzyme was inhibited under iron-limited growth conditions as well as by protease inhibitors. The AMPD of C. albicans showed a moderate increase in activity when cultures were grown in the presence of the divalent cations Mg2+, Ca2+, and Zn2+. Moreover, ADP, ATP, adenine, adenosine, deoxyribose and hypoxanthine increased the enzyme activity. Cultures grown in trypticase soy broth exhibited maximum AMPD activity compared with those grown in Sabouraud dextrose broth or Lee's medium.  相似文献   

13.
14.
Previous studies showed an association of the common functional polymorphism (C34T, Gln12Stop) in the adenosine monophosphate deaminase-1 (AMPD1) gene with survival in heart failure (HF) and/or coronary artery disease (CAD). The aim of the study was to search for other mutations in selected regions of the AMPD1 gene in Polish CAD and HF patients, and to analyze their associations with obesity and diabetes. Exons 2, 3, 5, and 7 of AMPD1 were scanned for mutations in 97 patients with CAD without HF (CAD+ HF-), 104 patients with HF (HF+), and 200 newborns from North-Western Poland using denaturing high-performance liquid chromatography (DHPLC), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and direct sequencing. Frequencies of AMPD1 C34T mutation, as well as novel A99G, G512A, IVS4-6delT, and C784T sequence alterations, were similar in the three groups, but 860T mutated allele was less frequent in the combined CAD+ HF- and HF+ groups than in the controls (1.7% vs. 4.3%, p=0.040). Heterozygous 34CT genotype was associated with lower (odds ratio [OR]=0.32, 95% confidence interval [CI]?=0.13-0.81) and 860AT with higher (OR=13.7, 95%CI=1.6-118) prevalence of diabetes or hyperglycemia in relation to wild-type homozygotes. Abdominal obesity was more frequent in 860AT patients than in wild-type homozygotes and 34CT heterozygotes (86% vs. 40% vs. 29%, p<0.05). Nine genes containing polymorphisms linked with AMPD1 C34T mutation were found in the HapMap database. AMPD1 C34T nonsense mutation is associated with reduced prevalence of diabetes and obesity in patients with CAD or HF, but A860T substitution seems to exert opposite metabolic effects and should always be accounted for in the studies of the AMPD1 genotype.  相似文献   

15.
Alternative splicing of the 12-base exon 2 of the adenosine monophosphate deaminase (AMPD) gene is subject to regulation by both cis- and trans-regulatory signals. The extent of exon 2 inclusion is stage- and cell type-specific and is subject to the physiological state of the cell. In adult skeletal muscle, a cell type that regulates the activity of this allosteric enzyme at several levels, the exon 2-plus form of AMPD, predominates. We have performed a systematic analysis of the cis-acting regulatory sequences that reside in the intron immediately downstream of this mini-exon. A complex element comprising sequences that enhance exon 2 inclusion and sequences that counteract this effect resides in the middle of this intron. We demonstrate that the enhancing component is bipartite, with more than a kilobase of sequence separating the two functional sites. The presence of even minimal levels the mini-exon in the fully processed AMPD mRNA requires both of these sites, neither of which appears in any other published splicing enhancer. An RNA binding activity derived from a muscle cell line requires both of the enhancing sites. Mutations in either of the sites that eliminate exon 2 inclusion abrogate this binding activity.  相似文献   

16.
Similar symptoms observed in Myasthenia gravis (MG) can be also detected in the case of skeletal muscle AMP-deaminase deficiency. We compared the activity and expression of AMP-deaminase (AMPD) products in skeletal muscles of MG patients and MG-free individuals. The activity of AMP-deaminase in the muscles of MG patients was significantly higher than in the controls and was 2.05 µmol/min/mg protein (±0.31). The two groups differ in level of AMPD product expression. Furthermore in MG-group molecular size of isoform AMPD1 is 90 kDa in contrast to MG-free group where is present 70 kDa isoform of enzyme. The data suggests that the disturbances in transmission of neuronal signaling, taking place in the skeletal muscles of MG patients, may also change energetic metabolism of the affected muscles by changing molecular mass of isoform.  相似文献   

17.
4-Pyridone-3-carboxamide-1-beta-D-ribonucleoside (4PYR) is an endogenously produced nucleoside that has recently been identified as a substrate for intracellular phosphorylation to form nucleotide derivatives. Low level of 4PYR is normally present in human plasma, but 4PYR massively accumulates in patients with renal failure. This study aimed to evaluate effects of 4PYR and its monophosphate derivative (4PYMP) on several enzymes of nucleotide metabolism in homogenates and intact cells. Activities of adenosine monophosphate deaminase (AMPD), adenosine deaminase, ecto-5′-nucleotidase (e5NT), adenine phosphoribosyltransferase (APRT), hypoxanthine/guanine phosphoribosyltransferase, purine nucleoside phosphorylase, and S-adenosylhomocysteine hydrolase (SAHH) were evaluated in erythrocyte lysates, rat heart homogenates, and in the intact rat cardiomyocytes by high performance liquid chromatography–based assays. 4PYMP caused significant inhibition of AMPD in both erythrocyte lysate and heart homogenate with 50% inhibitory concentration (IC50) of 74 and 55 μM, respectively. Inhibition of e5NT in heart homogenates was also noted with IC50 of 63 μM. 4PYMP slightly inhibited APRT and 4PYR caused moderate activation of SAHH. No effects on other enzymes studied were noted. Inhibition of AMPD by 4PYMP in homogenates was confirmed in the intact cell experiments with isolated cardiomyocytes that were allowed to accumulate 4PYMP by incubation with 4PYR. We conclude that among pathways studied, most important is the effect of 4PYMP on AMPD and that such effect could be one of the consequences of elevated plasma 4PYR concentration.  相似文献   

18.
Fatty liver (hepatic steatosis) is associated with nucleotide turnover, loss of ATP and generation of adenosine monophosphate (AMP). It is well known that in fatty liver, activity of the AMP-activated kinase (AMPK) is reduced and that its stimulation can prevent hepatic steatosis by both enhancing fat oxidation and reducing lipogenesis. Here we show that another AMP dependent enzyme, AMPD2, has opposing effects on fatty acid oxidation when compared to AMPK. In human hepatocytres, AMPD2 activation –either by overexpression or by lowering intracellular phosphate levels with fructose- is associated with a significant reduction in AMPK activity. Likewise, silencing of AMPK spontaneously increases AMPD activity, demonstrating that these enzymes counter-regulate each other. Furthermore, we show that a downstream product of AMP metabolism through AMPD2, uric acid, can inhibit AMPK activity in human hepatocytes. Finally, we show that fructose-induced fat accumulation in hepatocytes is due to a dominant stimulation of AMPD2 despite stimulating AMPK. In this regard, AMPD2-deficient hepatocytes demonstrate a further activation of AMPK after fructose exposure in association with increased fatty acid oxidation, and conversely silencing AMPK enhances AMPD-dependent fat accumulation. In vivo, we show that sucrose fed rats also develop fatty liver that is blocked by metformin in association with both a reduction in AMPD activity and an increase in AMPK activity. In summary, AMPD and AMPK are both important in hepatic fat accumulation and counter-regulate each other. We present the novel finding that uric acid inhibits AMPK kinase activity in fructose-fed hepatocytes thus providing new insights into the pathogenesis of fatty liver.  相似文献   

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Background

Current treatments for idiopathic inflammatory myopathies (collectively called myositis) focus on the suppression of an autoimmune inflammatory response within the skeletal muscle. However, it has been observed that there is a poor correlation between the successful suppression of muscle inflammation and an improvement in muscle function. Some evidence in the literature suggests that metabolic abnormalities in the skeletal muscle underlie the weakness that continues despite successful immunosuppression. We have previously shown that decreased expression of a purine nucleotide cycle enzyme, adenosine monophosphate deaminase (AMPD1), leads to muscle weakness in a mouse model of myositis and may provide a mechanistic basis for muscle weakness. One of the downstream metabolites of this pathway, D-ribose, has been reported to alleviate symptoms of myalgia in patients with a congenital loss of AMPD1. Therefore, we hypothesized that supplementing exogenous D-ribose would improve muscle function in the mouse model of myositis. We treated normal and myositis mice with daily doses of D-ribose (4 mg/kg) over a 6-week time period and assessed its effects using a battery of behavioral, functional, histological and molecular measures.

Results

Treatment with D-ribose was found to have no statistically significant effects on body weight, grip strength, open field behavioral activity, maximal and specific forces of EDL, soleus muscles, or histological features. Histological and gene expression analysis indicated that muscle tissues remained inflamed despite treatment. Gene expression analysis also suggested that low levels of the ribokinase enzyme in the skeletal muscle might prevent skeletal muscle tissue from effectively utilizing D-ribose.

Conclusions

Treatment with daily oral doses of D-ribose showed no significant effect on either disease progression or muscle function in the mouse model of myositis.  相似文献   

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