Genome editing is a valuable tool to target specific DNA sequences for mutagenesis in the genomes of microbes, plants, and animals. Although different genome editing technologies are available, the clustered regularly...Genome editing is a valuable tool to target specific DNA sequences for mutagenesis in the genomes of microbes, plants, and animals. Although different genome editing technologies are available, the clustered regularly interspaced short palindromic repeats/Cas9 （CRISPR/ Cas9） system, which utilizes engineered endonucleases to generate a double-stranded DNA break （DSB） in the target DNA region and subsequently stimulates site-specific mutagenesis through DNA repair machineries, is emerging as a powerful genome editing tool for elucidating mecha- nisms of protection from plant viruses, plant disease resistance, and gene functions in basic and applied research. In this review, we provide an overview of recent advances in the CRISPR system associated genome editing in plants by focusing on application of this technology in model plants, crop plants, fruit plants, woody plants and grasses and discuss how genome editing associated with the CRISPR system can provide insights into genome modifications and functional genomics in plants.展开更多
In this study, we analyzed complete mtDNA sequences variation and genetic relationship among taurine, indicine and Bison groups. In total, 107 sequences from different breeds, 45 European (45 Italian), 16 Middle East ...In this study, we analyzed complete mtDNA sequences variation and genetic relationship among taurine, indicine and Bison groups. In total, 107 sequences from different breeds, 45 European (45 Italian), 16 Middle East Asian (seven Iranian and nine Iraqi), 41 Northeast Asian (34 Korean and seven Japanese), two Nellore (Bos indicus) and two American Bison bison (Ame. bison) were obtained from Gen-Bank database. One Korean Hanwoo (Bos taurus) sequence was generated using the SOLiDTM System. In total, 1370 polymorphic sites, representing 8.39% of the complete 107 mtDNA sequences (16,338 bp) were detected and of these, 1186 parsimony informative polymorphic sites were identified. Neighbor-joining tree indicated that Korean, Japanese, Iranian, Iraqi, and Italian cattle were closely related to one another, but are separated from B. Bison. The B. taurus mtDNA polymorphism was greater in the D-loop than in the other regions. The ATP8, ND3, ND5, and ND6 regions were also quite parsimony informative, similar to Cyt b. In addition, this study revealed a distinct genetic difference between Korean cattle and B. indicus.展开更多
基于SYBR Green Ⅰ荧光染料与双链DNA（dsDNA）结合产生荧光的原理,建立一种高精度、高通量的双链DNA定量方法。将梯度稀释后的基因组DNA及已知浓度的λDNA与等体积的SYBR Green Ⅰ（4×）充分混合后,利用荧光定量PCR仪采集荧光信号...基于SYBR Green Ⅰ荧光染料与双链DNA（dsDNA）结合产生荧光的原理,建立一种高精度、高通量的双链DNA定量方法。将梯度稀释后的基因组DNA及已知浓度的λDNA与等体积的SYBR Green Ⅰ（4×）充分混合后,利用荧光定量PCR仪采集荧光信号,以ROX（1×）作为校正染料进行定量分析;同时利用紫外分光光度计对样品进行平行测定,比较该方法与紫外分光光度法的检测限与准确度。紫外分光光度法的检测限为2ng/μl,而SYBR Green Ⅰ荧光定量法的检测限可达到0.015ng/μl,并且在0.015-2ng/μl范围内,SYBR Green Ⅰ荧光强度与λDNA浓度呈线性关系（R2=0.9999）,比紫外分光光度法灵敏100倍以上,并可准确定量低纯度的DNA样品。此方法具有重复性好、高通量的特点,仅需少量的生物样本即可满足定量要求,为分子生物学研究及临床检验等多个领域提供了一种可靠的dsDNA定量方法。展开更多
目的优化试验条件,建立用于 DNA 甲基化组测定的限制性酶切扫描技术(RLGS)。方法选取冰冻胃癌组织及其周围非癌组织各2份,提取基因组大分子 DNA(>50 000 bp),用甲基化敏感的限制性内切酶 Not Ⅰ等对 DNA 进行多重酶切、同位素^(32)P...目的优化试验条件,建立用于 DNA 甲基化组测定的限制性酶切扫描技术(RLGS)。方法选取冰冻胃癌组织及其周围非癌组织各2份,提取基因组大分子 DNA(>50 000 bp),用甲基化敏感的限制性内切酶 Not Ⅰ等对 DNA 进行多重酶切、同位素^(32)P 标记、二维电泳、扫描分析,并且根据已有的位点 DNA 序列数据库,确定所得扫描图谱上位点所对应的序列信息。结果成功得到RLGS 扫描图;有效点平均在1200个左右,标本质量较好的图谱平均可获得有效点1800个左右,与国外实验室的平均水平相当,经过比对可找出放射自显影信号强度减弱或增强的点,结果可重复,并能在 Not Ⅰ-EcoRV 克隆文库中找到这些点所对应的序列信息。结论成功建立了 RLGS 技术平台,并能够稳定工作。展开更多
The DNA damage response（DDR） is a complex biological system activated by different types of DNA damage.Mutations in certain components of the DDR machinery can lead to genomic instability disorders that culminate in...The DNA damage response（DDR） is a complex biological system activated by different types of DNA damage.Mutations in certain components of the DDR machinery can lead to genomic instability disorders that culminate in tissue degeneration,premature aging,and various types of cancers.Intriguingly,malfunctioning DDR plays a role in the etiology of late onset brain degenerative disorders such as Parkinson＇s,Alzheimer＇s,and Huntington＇s diseases.For many years,brain degenerative disorders were thought to result from aberrant neural death.Here we discuss the evidence that supports our novel hypothesis that brain degenerative diseases involve dysfunction of glial cells（astrocytes,microglia,and oligodendrocytes）.Impairment in the functionality of glial cells results in pathological neuro-glial interactions that,in turn,generate a ‘‘hostile＂ environment that impairs the functionality of neuronal cells.These events can lead to systematic neural demise on a scale that appears to be proportional to the severity of the neurological deficit.展开更多
文摘Genome editing is a valuable tool to target specific DNA sequences for mutagenesis in the genomes of microbes, plants, and animals. Although different genome editing technologies are available, the clustered regularly interspaced short palindromic repeats/Cas9 （CRISPR/ Cas9） system, which utilizes engineered endonucleases to generate a double-stranded DNA break （DSB） in the target DNA region and subsequently stimulates site-specific mutagenesis through DNA repair machineries, is emerging as a powerful genome editing tool for elucidating mecha- nisms of protection from plant viruses, plant disease resistance, and gene functions in basic and applied research. In this review, we provide an overview of recent advances in the CRISPR system associated genome editing in plants by focusing on application of this technology in model plants, crop plants, fruit plants, woody plants and grasses and discuss how genome editing associated with the CRISPR system can provide insights into genome modifications and functional genomics in plants.
文摘In this study, we analyzed complete mtDNA sequences variation and genetic relationship among taurine, indicine and Bison groups. In total, 107 sequences from different breeds, 45 European (45 Italian), 16 Middle East Asian (seven Iranian and nine Iraqi), 41 Northeast Asian (34 Korean and seven Japanese), two Nellore (Bos indicus) and two American Bison bison (Ame. bison) were obtained from Gen-Bank database. One Korean Hanwoo (Bos taurus) sequence was generated using the SOLiDTM System. In total, 1370 polymorphic sites, representing 8.39% of the complete 107 mtDNA sequences (16,338 bp) were detected and of these, 1186 parsimony informative polymorphic sites were identified. Neighbor-joining tree indicated that Korean, Japanese, Iranian, Iraqi, and Italian cattle were closely related to one another, but are separated from B. Bison. The B. taurus mtDNA polymorphism was greater in the D-loop than in the other regions. The ATP8, ND3, ND5, and ND6 regions were also quite parsimony informative, similar to Cyt b. In addition, this study revealed a distinct genetic difference between Korean cattle and B. indicus.
文摘基于SYBR Green Ⅰ荧光染料与双链DNA（dsDNA）结合产生荧光的原理,建立一种高精度、高通量的双链DNA定量方法。将梯度稀释后的基因组DNA及已知浓度的λDNA与等体积的SYBR Green Ⅰ（4×）充分混合后,利用荧光定量PCR仪采集荧光信号,以ROX（1×）作为校正染料进行定量分析;同时利用紫外分光光度计对样品进行平行测定,比较该方法与紫外分光光度法的检测限与准确度。紫外分光光度法的检测限为2ng/μl,而SYBR Green Ⅰ荧光定量法的检测限可达到0.015ng/μl,并且在0.015-2ng/μl范围内,SYBR Green Ⅰ荧光强度与λDNA浓度呈线性关系（R2=0.9999）,比紫外分光光度法灵敏100倍以上,并可准确定量低纯度的DNA样品。此方法具有重复性好、高通量的特点,仅需少量的生物样本即可满足定量要求,为分子生物学研究及临床检验等多个领域提供了一种可靠的dsDNA定量方法。
文摘目的优化试验条件,建立用于 DNA 甲基化组测定的限制性酶切扫描技术(RLGS)。方法选取冰冻胃癌组织及其周围非癌组织各2份,提取基因组大分子 DNA(>50 000 bp),用甲基化敏感的限制性内切酶 Not Ⅰ等对 DNA 进行多重酶切、同位素^(32)P 标记、二维电泳、扫描分析,并且根据已有的位点 DNA 序列数据库,确定所得扫描图谱上位点所对应的序列信息。结果成功得到RLGS 扫描图;有效点平均在1200个左右,标本质量较好的图谱平均可获得有效点1800个左右,与国外实验室的平均水平相当,经过比对可找出放射自显影信号强度减弱或增强的点,结果可重复,并能在 Not Ⅰ-EcoRV 克隆文库中找到这些点所对应的序列信息。结论成功建立了 RLGS 技术平台,并能够稳定工作。
基金funded by the Israel Science Foundation (Grant Nos.rants 549/12 and 421/15)German Israeli Foundation (Grant No.I-192-418.13-2014)Joint ItalianIsraeli Laboratory on Application of Neuroscience (Grant No.590308)
文摘The DNA damage response（DDR） is a complex biological system activated by different types of DNA damage.Mutations in certain components of the DDR machinery can lead to genomic instability disorders that culminate in tissue degeneration,premature aging,and various types of cancers.Intriguingly,malfunctioning DDR plays a role in the etiology of late onset brain degenerative disorders such as Parkinson＇s,Alzheimer＇s,and Huntington＇s diseases.For many years,brain degenerative disorders were thought to result from aberrant neural death.Here we discuss the evidence that supports our novel hypothesis that brain degenerative diseases involve dysfunction of glial cells（astrocytes,microglia,and oligodendrocytes）.Impairment in the functionality of glial cells results in pathological neuro-glial interactions that,in turn,generate a ‘‘hostile＂ environment that impairs the functionality of neuronal cells.These events can lead to systematic neural demise on a scale that appears to be proportional to the severity of the neurological deficit.
