细胞质雄性不育(cytoplasmic male sterility,CMS)在植物杂交育种中广为应用,其发生与线粒体中的CMS基因有着密切关系。但是由于线粒体基因敲除或下调表达技术手段十分有限,各种作物中大量鉴定出的CMS基因都未能进行功能验证。拟南芥(Ar...细胞质雄性不育(cytoplasmic male sterility,CMS)在植物杂交育种中广为应用,其发生与线粒体中的CMS基因有着密切关系。但是由于线粒体基因敲除或下调表达技术手段十分有限,各种作物中大量鉴定出的CMS基因都未能进行功能验证。拟南芥(Arabidopsis thaliana)线粒体RPF(RNA processing factor)蛋白属于PPR(pentatricopeptide repeat)蛋白家族中的P亚类,由6~8个PPR重复基序组成。每个PPR基序含有35个氨基酸,其第5位和第35位氨基酸对于识别线粒体靶向RNA的一个碱基起到关键作用,由此建立了"PPR密码"规则。拟南芥RPF蛋白以序列识别的方式结合并加工线粒体RNA转录本的5′末端,但目前尚不清楚该加工过程的具体细节及其生理功能。本文综述了RPF蛋白编码基因RPF1~RPF7的鉴定及其功能分析,以及利用人工设计的RPF2蛋白使线粒体基因nad6下调表达的研究进展。植物线粒体RPF蛋白编码基因的鉴定及其功能和应用,对于阐明线粒体mRNA 5′末端加工的分子机制,以及利用反向遗传学手段研究线粒体基因功能,具有重要的科学意义和应用前景。展开更多
目的:构建大鼠线粒体转录因子TFAM(A)、线粒体转录因子TFB1M(B1)、线粒体转录TFB2M(B2)及线粒体RNA聚合酶(POLRMT)的质粒标准品,为检测内耳细胞线粒体TFAM、TFB1M、TFB2M、POLRMT的mRNA表达水平做基础。方法:设计特异性引物和探针,提取...目的:构建大鼠线粒体转录因子TFAM(A)、线粒体转录因子TFB1M(B1)、线粒体转录TFB2M(B2)及线粒体RNA聚合酶(POLRMT)的质粒标准品,为检测内耳细胞线粒体TFAM、TFB1M、TFB2M、POLRMT的mRNA表达水平做基础。方法:设计特异性引物和探针,提取大鼠内耳组织总mRNA逆转录成c DNA,PCR扩增、纯化目的片段,将纯化产物与p Zero Back/blunt载体重组,提取重组质粒,经测序鉴定后,用实时荧光绝对定量PCR建立标准曲线。结果:测序结果与各目的序列一致,获得良好的标准曲线(R2>0.99)。结论:成功构建了各目的基因的质粒标准品。展开更多
A 605 bp section of mitochondrial 16S rRNA gene from Paralichthys olivaceus, Pseudorhombus cinnamomeus, Psetta maxima and Kareius bicoloratus, which represent 3 families of Order Pleuronectiformes was amplified by PCR...A 605 bp section of mitochondrial 16S rRNA gene from Paralichthys olivaceus, Pseudorhombus cinnamomeus, Psetta maxima and Kareius bicoloratus, which represent 3 families of Order Pleuronectiformes was amplified by PCR and sequenced to show the molecular systematics of Pleuronectiformes for comparison with related gene sequences of other 6 flatfish downloaded from GenBank. Phylogenetic analysis based on ge- netic distance from related gene sequences of 10 flatfish showed that this method was ideal to explore the rela- tionship between species, genera and families. Phylogenetic trees set-up is based on neighbor-joining, maximum parsimony and maximum likelihood methods that accords to the general rule of Pleuronectiformes evolution. But they also resulted in some confusion. Unlike data from morphological characters, P. olivaceus clustered with K. bicoloratus, but P. cinnamomeus did not cluster with P. olivaceus, which is worth further studying.展开更多
AIM: To design a novel method to rapidly detect the quantitative alteration of mtRNA in patients with tumors.METHODS: Oligo 6.22 and Primer Premier 5.0 bio-soft were used to design 15 pairs of primers of mtRNA cDNA pr...AIM: To design a novel method to rapidly detect the quantitative alteration of mtRNA in patients with tumors.METHODS: Oligo 6.22 and Primer Premier 5.0 bio-soft were used to design 15 pairs of primers of mtRNA cDNA probes in light of the functional and structural property of mtDNA, and then RT-PCR amplification was used to produce 15 probes of mtRNA from one normal gastric mucosal tissue. Total RNA extracted from 9 gastric cancers and corresponding normal gastric mucosal tissues was reverse transcribed into cDNA labeled with fluorescein. The spotted mtDNA microarrays were made and hybridized. Finally,the microarrays were scanned with a GeneTACTM laser scanner to get the hybridized results. Northern blot was used to confirm the microarray results.RESULTS: The hybridized spots were distinct with clear and consistent backgrounds. After data was standardized according to the housekeeping genes, the results showed that the expression levels of some mitochondrial genes in gastric carcinoma were different from those in the corresponding non-cancerous regions.CONCLUSION: The mtDNA expression microarray can rapidly, massively and exactly detect the quantity of mtRNA in tissues and cells. In addition, the whole expressive information of mtRNA from a tumor patient on just one slide can be obtained using this method, providing an effective method to investigate the relationship between mtDNA expression and tumorigenesis.展开更多
RNase MRP RNA is the RNA subunit of the RNase mitochondrial RNA processing （MRP） enzyme complex that is involved in multiple cellular RNA processing events. Mutations on RNase MRP RNA gene （RMRP） cause a recessive...RNase MRP RNA is the RNA subunit of the RNase mitochondrial RNA processing （MRP） enzyme complex that is involved in multiple cellular RNA processing events. Mutations on RNase MRP RNA gene （RMRP） cause a recessively inherited developmental disorder, cartilage-hair hypoplasia （CHH）. The relationship of the genotype （RMRP mutation）, RNA processing deficiency of the RNase MRP complex, and the phenotype of CHH and other skeletal dysplasias is yet to be explored.展开更多
文摘目的:构建大鼠线粒体转录因子TFAM(A)、线粒体转录因子TFB1M(B1)、线粒体转录TFB2M(B2)及线粒体RNA聚合酶(POLRMT)的质粒标准品,为检测内耳细胞线粒体TFAM、TFB1M、TFB2M、POLRMT的mRNA表达水平做基础。方法:设计特异性引物和探针,提取大鼠内耳组织总mRNA逆转录成c DNA,PCR扩增、纯化目的片段,将纯化产物与p Zero Back/blunt载体重组,提取重组质粒,经测序鉴定后,用实时荧光绝对定量PCR建立标准曲线。结果:测序结果与各目的序列一致,获得良好的标准曲线(R2>0.99)。结论:成功构建了各目的基因的质粒标准品。
基金Supported by Natural Science Fund of China (No. 30271036) and Natural Science Fund of Shandong Province of China
文摘A 605 bp section of mitochondrial 16S rRNA gene from Paralichthys olivaceus, Pseudorhombus cinnamomeus, Psetta maxima and Kareius bicoloratus, which represent 3 families of Order Pleuronectiformes was amplified by PCR and sequenced to show the molecular systematics of Pleuronectiformes for comparison with related gene sequences of other 6 flatfish downloaded from GenBank. Phylogenetic analysis based on ge- netic distance from related gene sequences of 10 flatfish showed that this method was ideal to explore the rela- tionship between species, genera and families. Phylogenetic trees set-up is based on neighbor-joining, maximum parsimony and maximum likelihood methods that accords to the general rule of Pleuronectiformes evolution. But they also resulted in some confusion. Unlike data from morphological characters, P. olivaceus clustered with K. bicoloratus, but P. cinnamomeus did not cluster with P. olivaceus, which is worth further studying.
基金Supported by the National Natural Science Foundation of China,No.30371607
文摘AIM: To design a novel method to rapidly detect the quantitative alteration of mtRNA in patients with tumors.METHODS: Oligo 6.22 and Primer Premier 5.0 bio-soft were used to design 15 pairs of primers of mtRNA cDNA probes in light of the functional and structural property of mtDNA, and then RT-PCR amplification was used to produce 15 probes of mtRNA from one normal gastric mucosal tissue. Total RNA extracted from 9 gastric cancers and corresponding normal gastric mucosal tissues was reverse transcribed into cDNA labeled with fluorescein. The spotted mtDNA microarrays were made and hybridized. Finally,the microarrays were scanned with a GeneTACTM laser scanner to get the hybridized results. Northern blot was used to confirm the microarray results.RESULTS: The hybridized spots were distinct with clear and consistent backgrounds. After data was standardized according to the housekeeping genes, the results showed that the expression levels of some mitochondrial genes in gastric carcinoma were different from those in the corresponding non-cancerous regions.CONCLUSION: The mtDNA expression microarray can rapidly, massively and exactly detect the quantity of mtRNA in tissues and cells. In addition, the whole expressive information of mtRNA from a tumor patient on just one slide can be obtained using this method, providing an effective method to investigate the relationship between mtDNA expression and tumorigenesis.
文摘RNase MRP RNA is the RNA subunit of the RNase mitochondrial RNA processing （MRP） enzyme complex that is involved in multiple cellular RNA processing events. Mutations on RNase MRP RNA gene （RMRP） cause a recessively inherited developmental disorder, cartilage-hair hypoplasia （CHH）. The relationship of the genotype （RMRP mutation）, RNA processing deficiency of the RNase MRP complex, and the phenotype of CHH and other skeletal dysplasias is yet to be explored.
