Aim： To investigate the methylation status of the deleted in azoospermia 1（DAZ1） gene promoter region in different cell types. Methods： Using CpG island Searcher software, a CpG island was found in the promoter re...Aim： To investigate the methylation status of the deleted in azoospermia 1（DAZ1） gene promoter region in different cell types. Methods： Using CpG island Searcher software, a CpG island was found in the promoter region of the DAZ1 gene. The methylation status of this region was analyzed in sperm and leukocytes by bisulfited sequencing. Results： The methylation status of the CpG island in the DAZ1 gene promoter region differed in leukocytes and sperm： it was methylated in leukocytes, but unmethylated in sperm. Conclusion： A differentially methylated region of the DAZ1 gene exists in spermatic and somatic cells, suggesting that methylation of this region may regulate DAZ1 gene expression in different tissues. （Asian J Androl 2006 Jan; 8：61-67 ）展开更多
为克隆鸡Daz(lDeleted in azoospermia-like,Dazl)基因CDS序列,通过构建eGFP标记的DAZL真核表达载体,实现该基因产物的亚细胞定位以探究其生物信息学功能。提取1日龄鸡睾丸总RNA,通过巢式PCR方法扩增出Dazl基因的CDS,构建真核表达载体pE...为克隆鸡Daz(lDeleted in azoospermia-like,Dazl)基因CDS序列,通过构建eGFP标记的DAZL真核表达载体,实现该基因产物的亚细胞定位以探究其生物信息学功能。提取1日龄鸡睾丸总RNA,通过巢式PCR方法扩增出Dazl基因的CDS,构建真核表达载体pEGFP-C1-DAZL。采用LipofectaminTMLTX介导重组表达载体pEGFP-C1-DAZL转染CEF细胞,48h后于荧光倒置显微镜下观察其表达定位,同时利用RT-PCR和Western-blot进一步鉴定eGFP-Dazl和蛋白水平的表达情况。结果表明:克隆出Dazl基因的完整CDS,长度870bp,共编码289个氨基酸,与公布的鸡Dazl基因(GenBank登陆号:NM_204218)CDS区同源性达99%,编码氨基酸完全一致。酶切鉴定和序列分析均表明真核表达载体pEGFP-C1-DAZL构建成功。转染48h后,RT-PCR和Western-blot分别检测到949bp特异条带和59.7ku的融合蛋白。荧光显微镜观察显示,融合蛋白(eGFP-Dazl)主要分布于细胞核。展开更多
目的:探讨精子质量及DAZ(deleted in azoospermia)基因与男性不育症的关系。方法:应用计算机辅助精子质量检测系统进行检测精子质量;用PCR方法检测DAZ基因。结果:不育组(n=600)精子质量明显低于正常生育组(n=25)(精子密度不育组34.43...目的:探讨精子质量及DAZ(deleted in azoospermia)基因与男性不育症的关系。方法:应用计算机辅助精子质量检测系统进行检测精子质量;用PCR方法检测DAZ基因。结果:不育组(n=600)精子质量明显低于正常生育组(n=25)(精子密度不育组34.43±43.43×106/ml,生育组92.86±62.45×106/ml,P<0.01;活率分别为42.93±25.33%、75.25±12.75%,P<0.01;各级精子所占百分比相比也有显著性差异),男性不育患者中DAZ基因缺失占一定比例(占13.91%),无精子症所占比例较大达36.6%。结论:精液分析是男性不育患者最基本的诊断手段,精子质量差是男性不育最基本的原因;DAZ基因与精子的生成有密切关系,也可引起男性不育。展开更多
文摘Aim： To investigate the methylation status of the deleted in azoospermia 1（DAZ1） gene promoter region in different cell types. Methods： Using CpG island Searcher software, a CpG island was found in the promoter region of the DAZ1 gene. The methylation status of this region was analyzed in sperm and leukocytes by bisulfited sequencing. Results： The methylation status of the CpG island in the DAZ1 gene promoter region differed in leukocytes and sperm： it was methylated in leukocytes, but unmethylated in sperm. Conclusion： A differentially methylated region of the DAZ1 gene exists in spermatic and somatic cells, suggesting that methylation of this region may regulate DAZ1 gene expression in different tissues. （Asian J Androl 2006 Jan; 8：61-67 ）
文摘为克隆鸡Daz(lDeleted in azoospermia-like,Dazl)基因CDS序列,通过构建eGFP标记的DAZL真核表达载体,实现该基因产物的亚细胞定位以探究其生物信息学功能。提取1日龄鸡睾丸总RNA,通过巢式PCR方法扩增出Dazl基因的CDS,构建真核表达载体pEGFP-C1-DAZL。采用LipofectaminTMLTX介导重组表达载体pEGFP-C1-DAZL转染CEF细胞,48h后于荧光倒置显微镜下观察其表达定位,同时利用RT-PCR和Western-blot进一步鉴定eGFP-Dazl和蛋白水平的表达情况。结果表明:克隆出Dazl基因的完整CDS,长度870bp,共编码289个氨基酸,与公布的鸡Dazl基因(GenBank登陆号:NM_204218)CDS区同源性达99%,编码氨基酸完全一致。酶切鉴定和序列分析均表明真核表达载体pEGFP-C1-DAZL构建成功。转染48h后,RT-PCR和Western-blot分别检测到949bp特异条带和59.7ku的融合蛋白。荧光显微镜观察显示,融合蛋白(eGFP-Dazl)主要分布于细胞核。
文摘目的:探讨精子质量及DAZ(deleted in azoospermia)基因与男性不育症的关系。方法:应用计算机辅助精子质量检测系统进行检测精子质量;用PCR方法检测DAZ基因。结果:不育组(n=600)精子质量明显低于正常生育组(n=25)(精子密度不育组34.43±43.43×106/ml,生育组92.86±62.45×106/ml,P<0.01;活率分别为42.93±25.33%、75.25±12.75%,P<0.01;各级精子所占百分比相比也有显著性差异),男性不育患者中DAZ基因缺失占一定比例(占13.91%),无精子症所占比例较大达36.6%。结论:精液分析是男性不育患者最基本的诊断手段,精子质量差是男性不育最基本的原因;DAZ基因与精子的生成有密切关系,也可引起男性不育。
