为探究丝氨酸蛋白酶抑制剂分支A成员1(Serpin Peptidase Inhibitor Clade A Member 1,SERPINA1)基因多态性对绵羊肉质的影响,本试验选取33只8月龄的双乾肉羊(Shuangqian Mutton Sheep)作为研究对象,筛查SERPINA1基因第1外显子的单核苷...为探究丝氨酸蛋白酶抑制剂分支A成员1(Serpin Peptidase Inhibitor Clade A Member 1,SERPINA1)基因多态性对绵羊肉质的影响,本试验选取33只8月龄的双乾肉羊(Shuangqian Mutton Sheep)作为研究对象,筛查SERPINA1基因第1外显子的单核苷酸多态性(Single Nucleotide Polymorphism,SNP)位点,并对其肉质性状和脂肪酸含量进行测定,分析SNP与双乾肉羊肉质性状和脂肪酸含量间的关联性。结果表明:SERPINA1基因第1外显子第66位碱基存在C>T突变。该位点的遗传纯合度(Ho)为0.5900,有效等位基因数是1.6949,多态信息含量(PIC)为0.3260,处于Hardy-Weinberg平衡(P>0.05),属于中度多态。就肉质性状而言,该位点TT和CC基因型个体的柔嫩度显著高于TC基因型个体,而在脂肪酸含量方面,TC和CC基因型个体的棕榈油酸含量显著高于TT基因型个体。由此可见,绵羊SERPINA1基因的第1外显子C66T等位基因多态性与双乾肉羊肉嫩度和棕榈酸含量显著相关,该位点可成为预测并筛选具有高肉质性状和高脂肪酸含量的个体的重要SNP标记。展开更多
本研究旨在克隆西农萨能奶山羊SERPINA1基因的CDS区,采用生物软件和在线预测工具进行生物信息学分析,通过实时荧光定量PCR技术检测SERPINA1基因在西农萨能奶山羊各组织间mRNA的表达水平。根据GenBank中山羊SERPINA1基因CDS区序列(登录号...本研究旨在克隆西农萨能奶山羊SERPINA1基因的CDS区,采用生物软件和在线预测工具进行生物信息学分析,通过实时荧光定量PCR技术检测SERPINA1基因在西农萨能奶山羊各组织间mRNA的表达水平。根据GenBank中山羊SERPINA1基因CDS区序列(登录号:XM_018066209.1),利用Primer Premier 5.0软件设计特异性引物,RT-PCR扩增目的基因,构建原核表达载体测序后对序列进行生物信息学分析;采集西农萨能奶山羊心脏、肝脏、脾脏、肺脏、乳腺、肾脏、肌肉、瘤胃和小肠组织,提取组织RNA,反转录为cDNA模板,设计特异性定量引物,进行实时荧光定量PCR,检测SERPINA1基因在不同组织中的表达差异。结果显示,西农萨能奶山羊SERPINA1基因CDS区全长1326 bp,编码441个氨基酸;同源性比对分析显示,西农萨能奶山羊与山羊、绵羊、牛和小鼠SERPINA1基因核苷酸序列同源性分别为100%、98.6%、95.7%和71.6%,与山羊亲缘关系最近,其次是绵羊。SERPINA1蛋白分子质量为48.71 ku,等电点为5.71,为跨膜亲水蛋白;SERPINA1氨基酸序列分别有62个磷酸化位点,3个跨膜区结构。组织表达分析显示,SERPINA1基因在西农萨能奶山羊肝脏组织中显著高表达(P<0.05),其次是乳腺组织,在肺脏组织中表达量最低。研究结果为进一步探究SERPINA1基因在奶山羊乳蛋白合成代谢中的作用提供理论依据。展开更多
AIM:To determine the frequencies of mutations that cause inherited monogenic liver disorders in patients with chronic hepatitis C. METHODS:This study included 86 patients with chronic hepatitis C(55 men, 31 women; mea...AIM:To determine the frequencies of mutations that cause inherited monogenic liver disorders in patients with chronic hepatitis C. METHODS:This study included 86 patients with chronic hepatitis C(55 men, 31 women; mean age at diagnosis, 38.36 ± 14.52 years) who had undergone antiviral therapy comprising pegylated interferon and ribavirin. Viral load, biochemical parameter changes, and liver biopsy morphological data were evaluated in all patients. The control group comprised 271 unrelated individuals representing the general population of Latvia for mutation frequency calculations. The most frequent mutations that cause inherited liver disorders [gene(mutation): ATP7B(H1069Q), HFE(C282Y, H63D),UGT1A1(TA)7, and SERPINA1(PiZ)] were detected by polymerase chain reaction(PCR), bidirectional PCR allele-specific amplification, restriction fragment length polymorphism analysis, and sequencing. RESULTS: The viral genotype was detected in 80 of the 86 patients. Viral genotypes 1, 2, and 3 were present in 61(76%), 7(9%), and 12(15%) patients, respectively. Among all 86 patients, 50(58%) reached an early viral response and 70(81%) reached a sustained viral response. All 16 patients who did not reach a sustained viral response had viral genotype 1. Casecontrol analysis revealed a statistically significant difference in only the H1069Q mutation between patients and controls(patients, 0.057; controls, 0.012; odds ratio, 5.514; 95%CI: 1.119-29.827, P = 0.022). However, the H1069Q mutation was not associated with antiviral treatment outcomes or biochemical indices. The(TA) 7 mutation of the UGT1A1 gene was associated with decreased ferritin levels(beta regression coefficient =-295.7, P = 0.0087). CONCLUSION: Genetic mutations that cause inherited liver diseases in patients with hepatitis C should be studied in detail.展开更多
文摘本研究旨在克隆西农萨能奶山羊SERPINA1基因的CDS区,采用生物软件和在线预测工具进行生物信息学分析,通过实时荧光定量PCR技术检测SERPINA1基因在西农萨能奶山羊各组织间mRNA的表达水平。根据GenBank中山羊SERPINA1基因CDS区序列(登录号:XM_018066209.1),利用Primer Premier 5.0软件设计特异性引物,RT-PCR扩增目的基因,构建原核表达载体测序后对序列进行生物信息学分析;采集西农萨能奶山羊心脏、肝脏、脾脏、肺脏、乳腺、肾脏、肌肉、瘤胃和小肠组织,提取组织RNA,反转录为cDNA模板,设计特异性定量引物,进行实时荧光定量PCR,检测SERPINA1基因在不同组织中的表达差异。结果显示,西农萨能奶山羊SERPINA1基因CDS区全长1326 bp,编码441个氨基酸;同源性比对分析显示,西农萨能奶山羊与山羊、绵羊、牛和小鼠SERPINA1基因核苷酸序列同源性分别为100%、98.6%、95.7%和71.6%,与山羊亲缘关系最近,其次是绵羊。SERPINA1蛋白分子质量为48.71 ku,等电点为5.71,为跨膜亲水蛋白;SERPINA1氨基酸序列分别有62个磷酸化位点,3个跨膜区结构。组织表达分析显示,SERPINA1基因在西农萨能奶山羊肝脏组织中显著高表达(P<0.05),其次是乳腺组织,在肺脏组织中表达量最低。研究结果为进一步探究SERPINA1基因在奶山羊乳蛋白合成代谢中的作用提供理论依据。
基金Supported by The Latvian Council of Science,National Project,Nos.09.1384 and 10.0010.02
文摘AIM:To determine the frequencies of mutations that cause inherited monogenic liver disorders in patients with chronic hepatitis C. METHODS:This study included 86 patients with chronic hepatitis C(55 men, 31 women; mean age at diagnosis, 38.36 ± 14.52 years) who had undergone antiviral therapy comprising pegylated interferon and ribavirin. Viral load, biochemical parameter changes, and liver biopsy morphological data were evaluated in all patients. The control group comprised 271 unrelated individuals representing the general population of Latvia for mutation frequency calculations. The most frequent mutations that cause inherited liver disorders [gene(mutation): ATP7B(H1069Q), HFE(C282Y, H63D),UGT1A1(TA)7, and SERPINA1(PiZ)] were detected by polymerase chain reaction(PCR), bidirectional PCR allele-specific amplification, restriction fragment length polymorphism analysis, and sequencing. RESULTS: The viral genotype was detected in 80 of the 86 patients. Viral genotypes 1, 2, and 3 were present in 61(76%), 7(9%), and 12(15%) patients, respectively. Among all 86 patients, 50(58%) reached an early viral response and 70(81%) reached a sustained viral response. All 16 patients who did not reach a sustained viral response had viral genotype 1. Casecontrol analysis revealed a statistically significant difference in only the H1069Q mutation between patients and controls(patients, 0.057; controls, 0.012; odds ratio, 5.514; 95%CI: 1.119-29.827, P = 0.022). However, the H1069Q mutation was not associated with antiviral treatment outcomes or biochemical indices. The(TA) 7 mutation of the UGT1A1 gene was associated with decreased ferritin levels(beta regression coefficient =-295.7, P = 0.0087). CONCLUSION: Genetic mutations that cause inherited liver diseases in patients with hepatitis C should be studied in detail.
