荧光原位杂交技术(fluorescence in situ hybridization,FISH)是植物分子细胞遗传学研究最为重要的手段之一。近些年,基于参考基因组设计的低拷贝寡聚核苷酸探针在FISH中应用得越来越广泛。然而,由于植物基因组中分布大量的重复序列,这...荧光原位杂交技术(fluorescence in situ hybridization,FISH)是植物分子细胞遗传学研究最为重要的手段之一。近些年,基于参考基因组设计的低拷贝寡聚核苷酸探针在FISH中应用得越来越广泛。然而,由于植物基因组中分布大量的重复序列,这使得oligo-FISH的分辨率存在一定局限性。利用包含多个荧光基团的荧光PCR引物,扩增出甘蔗染色体特异oligo探针,并进一步优化甘蔗的荧光原位杂交体系,提高了甘蔗oligo探针识别近缘物种染色体的效率。通过开发多荧光标记的甘蔗oligo探针以及甘蔗荧光杂交体系的优化,有效拓宽荧光信号的最小分辨率,提高信噪比(signal-to-noise ratio,SNR),并成功基于甘蔗oligo探针对高粱1-10号染色体分型。多荧光标记引物增强oligo探针信号的新方法及FISH体系的优化为今后在其他物种中提高oligo-FISH鉴定染色体及捕捉微弱的荧光信号提供了参考。展开更多
[目的]本文旨在开发纤毛鹅观草特异的寡核苷酸(oligonucleotide,oligo)探针,进一步完善纤毛鹅观草染色体鉴定技术。[方法]利用前期选育的普通小麦-纤毛鹅观草异附加系DA2S^(c)L和DA6S^(c),通过流式分拣和基因组二代测序获得纤毛鹅观草...[目的]本文旨在开发纤毛鹅观草特异的寡核苷酸(oligonucleotide,oligo)探针,进一步完善纤毛鹅观草染色体鉴定技术。[方法]利用前期选育的普通小麦-纤毛鹅观草异附加系DA2S^(c)L和DA6S^(c),通过流式分拣和基因组二代测序获得纤毛鹅观草染色体6S^(c)和染色体臂2S^(c)长臂的基因组序列,利用Repeatexplorer2/TAREAN软件从中鉴定出卫星重复序列并设计成oligo探针,通过荧光原位杂交(fluorescence in situ hybridization,FISH)分析所开发oligo探针的应用价值。[结果]从6S^(c)和2S^(c)L基因组序列中共鉴定出11个卫星重复序列并开发成11个oligo探针,oligo-FISH结果表明,其中6个探针可以在纤毛鹅观草染色体上产生明显杂交信号,而在小麦染色体上未产生明显杂交信号,可用于特异鉴定小麦背景中的纤毛鹅观草染色体或片段。对1套普通小麦-纤毛鹅观草异附加系oligo-FISH分析发现,oligo-2S^(c)L^(-1)63和oligo-6S^(c)-111仅在S^(c)基因组染色体上产生明显信号,可作为纤毛鹅观草S^(c)组特异探针;将oligo-2S^(c)L-161和oligo-2S^(c)L-163组合,对1整套二体异附加系进行双色oligo-FISH,构建了纤毛鹅观草染色体的oligo-FISH核型。[结论]本研究提供了纤毛鹅观草重复序列组成的初步信息,开发的探针能特异鉴定纤毛鹅观草染色体,阐明纤毛鹅观草2个基因组的来源和分化,对推动纤毛鹅观草优异基因的转移和利用有重要意义。展开更多
In the present study, we examine whether selected genetic polymorphisms cont ribute to the development of cerebral palsy (CP) in very preterm infants. Subjec ts were 96 singleton infants with later- diagnosed CP and 1...In the present study, we examine whether selected genetic polymorphisms cont ribute to the development of cerebral palsy (CP) in very preterm infants. Subjec ts were 96 singleton infants with later- diagnosed CP and 119 control children, white non- Hispanic (n for CP = 74, controls = 88) or white Hispanic (CP = 22, controls = 31), born < 32 wk gestation. Presence of CP was identified through state service agencies, with review of medical records. DNA extracted from archi ved neonatal blood was genotyped using multi- locus polymerase chain reaction a mplification and immobilized sequence- specific oligonucleotide probes. Single nucleotide polymorphisms (SNPs) showing evidence of association with development of CP were endothelial nitric oxide synthase (eNOS) A(- 922)G, factor 7 (F7) a rg353gln and del(- 323)10bp- ins, and lymphotoxin A (LTA) thr26asn. In white n on- Hispanic children, beta- 2 adrenergic receptor gln27glu was associated wit h CP risk; in Hispanic children, plasminogen activator inhibitor- 1 (PAI- 1)- 4G(- 675)5G and G11053T were associated with risk of CP. In a logistic regress ion considering these SNPs simultaneously in non- Hispanics, an association wit h CP was observed for heterozygotes of eNOS - 922 (OR 3.0, CI 1.4- 6.4), F7 (O R 2.7, CI 1.1- 6.5), LTA (OR 2.1, CI 1.0- 4.6), and PAI- 1 (OR 3.2, CI 1.2- 8.7). Factor 5, Factor 2, methylene tetrahydrofolate reductase, tumor necrosis f actor- alpha, and other SNPs tested were not significantly associated with CP r isk. We conclude that further study of genetic factors that may influence suscep tibility to CP in very preterm infants is warranted.展开更多
文摘荧光原位杂交技术(fluorescence in situ hybridization,FISH)是植物分子细胞遗传学研究最为重要的手段之一。近些年,基于参考基因组设计的低拷贝寡聚核苷酸探针在FISH中应用得越来越广泛。然而,由于植物基因组中分布大量的重复序列,这使得oligo-FISH的分辨率存在一定局限性。利用包含多个荧光基团的荧光PCR引物,扩增出甘蔗染色体特异oligo探针,并进一步优化甘蔗的荧光原位杂交体系,提高了甘蔗oligo探针识别近缘物种染色体的效率。通过开发多荧光标记的甘蔗oligo探针以及甘蔗荧光杂交体系的优化,有效拓宽荧光信号的最小分辨率,提高信噪比(signal-to-noise ratio,SNR),并成功基于甘蔗oligo探针对高粱1-10号染色体分型。多荧光标记引物增强oligo探针信号的新方法及FISH体系的优化为今后在其他物种中提高oligo-FISH鉴定染色体及捕捉微弱的荧光信号提供了参考。
文摘[目的]本文旨在开发纤毛鹅观草特异的寡核苷酸(oligonucleotide,oligo)探针,进一步完善纤毛鹅观草染色体鉴定技术。[方法]利用前期选育的普通小麦-纤毛鹅观草异附加系DA2S^(c)L和DA6S^(c),通过流式分拣和基因组二代测序获得纤毛鹅观草染色体6S^(c)和染色体臂2S^(c)长臂的基因组序列,利用Repeatexplorer2/TAREAN软件从中鉴定出卫星重复序列并设计成oligo探针,通过荧光原位杂交(fluorescence in situ hybridization,FISH)分析所开发oligo探针的应用价值。[结果]从6S^(c)和2S^(c)L基因组序列中共鉴定出11个卫星重复序列并开发成11个oligo探针,oligo-FISH结果表明,其中6个探针可以在纤毛鹅观草染色体上产生明显杂交信号,而在小麦染色体上未产生明显杂交信号,可用于特异鉴定小麦背景中的纤毛鹅观草染色体或片段。对1套普通小麦-纤毛鹅观草异附加系oligo-FISH分析发现,oligo-2S^(c)L^(-1)63和oligo-6S^(c)-111仅在S^(c)基因组染色体上产生明显信号,可作为纤毛鹅观草S^(c)组特异探针;将oligo-2S^(c)L-161和oligo-2S^(c)L-163组合,对1整套二体异附加系进行双色oligo-FISH,构建了纤毛鹅观草染色体的oligo-FISH核型。[结论]本研究提供了纤毛鹅观草重复序列组成的初步信息,开发的探针能特异鉴定纤毛鹅观草染色体,阐明纤毛鹅观草2个基因组的来源和分化,对推动纤毛鹅观草优异基因的转移和利用有重要意义。
文摘In the present study, we examine whether selected genetic polymorphisms cont ribute to the development of cerebral palsy (CP) in very preterm infants. Subjec ts were 96 singleton infants with later- diagnosed CP and 119 control children, white non- Hispanic (n for CP = 74, controls = 88) or white Hispanic (CP = 22, controls = 31), born < 32 wk gestation. Presence of CP was identified through state service agencies, with review of medical records. DNA extracted from archi ved neonatal blood was genotyped using multi- locus polymerase chain reaction a mplification and immobilized sequence- specific oligonucleotide probes. Single nucleotide polymorphisms (SNPs) showing evidence of association with development of CP were endothelial nitric oxide synthase (eNOS) A(- 922)G, factor 7 (F7) a rg353gln and del(- 323)10bp- ins, and lymphotoxin A (LTA) thr26asn. In white n on- Hispanic children, beta- 2 adrenergic receptor gln27glu was associated wit h CP risk; in Hispanic children, plasminogen activator inhibitor- 1 (PAI- 1)- 4G(- 675)5G and G11053T were associated with risk of CP. In a logistic regress ion considering these SNPs simultaneously in non- Hispanics, an association wit h CP was observed for heterozygotes of eNOS - 922 (OR 3.0, CI 1.4- 6.4), F7 (O R 2.7, CI 1.1- 6.5), LTA (OR 2.1, CI 1.0- 4.6), and PAI- 1 (OR 3.2, CI 1.2- 8.7). Factor 5, Factor 2, methylene tetrahydrofolate reductase, tumor necrosis f actor- alpha, and other SNPs tested were not significantly associated with CP r isk. We conclude that further study of genetic factors that may influence suscep tibility to CP in very preterm infants is warranted.
