通过生物弹道转化和同源重组对酿酒酵母线粒体DNA进行定向改造。
Directed alteration of Saccharomyces cerevisiae mitochondrial DNA by biolistic transformation and homologous recombination.
作者信息
Bonnefoy Nathalie, Fox Thomas D
机构信息
Centre de Génétique Moléculaire, UPR 2167, Gif-sur-Yvette, France.
出版信息
Methods Mol Biol. 2007;372:153-66. doi: 10.1007/978-1-59745-365-3_11.
Abstract
Saccharomyces cerevisiae is currently the only species in which genetic transformation of mitochondria can be used to generate a wide variety of defined alterations in mitochondrial deoxyribonucleic acid (mtDNA). DNA sequences can be delivered into yeast mitochondria by microprojectile bombardment (biolistic transformation) and subsequently incorporated into mtDNA by the highly active homologous recombination machinery present in the organelle. Although transformation frequencies are relatively low, the availability of strong mitochondrial selectable markers for the yeast system, both natural and synthetic, makes the isolation of transformants routine. The strategies and procedures reviewed here allow the researcher to insert defined mutations into endogenous mitochondrial genes and to insert new genes into mtDNA. These methods provide powerful in vivo tools for the study of mitochondrial biology.
摘要
酿酒酵母是目前唯一一种可利用线粒体基因转化在其线粒体脱氧核糖核酸（mtDNA）中产生多种特定改变的物种。DNA序列可通过微粒轰击（生物弹道转化）导入酵母线粒体，随后通过该细胞器中存在的高活性同源重组机制整合到mtDNA中。尽管转化频率相对较低，但酵母系统中天然和合成的强大线粒体选择标记的可用性使得转化子的分离成为常规操作。本文综述的策略和程序使研究人员能够将特定突变插入内源性线粒体基因，并将新基因插入mtDNA。这些方法为线粒体生物学研究提供了强大的体内工具。
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本文引用的文献
1
Genetic transformation of Saccharomyces cerevisiae mitochondria.酿酒酵母线粒体的遗传转化
Methods Cell Biol. 2001;65:381-96. doi: 10.1016/s0091-679x(01)65022-2.
2
Mitochondrial translation of Saccharomyces cerevisiae COX2 mRNA is controlled by the nucleotide sequence specifying the pre-Cox2p leader peptide.酿酒酵母COX2 mRNA的线粒体翻译受指定前Cox2p前导肽的核苷酸序列控制。
Mol Cell Biol. 2001 Apr;21(7):2359-72. doi: 10.1128/MCB.21.7.2359-2372.2001.
3
In vivo analysis of mutated initiation codons in the mitochondrial COX2 gene of Saccharomyces cerevisiae fused to the reporter gene ARG8m reveals lack of downstream reinitiation.对酿酒酵母线粒体COX2基因中与报告基因ARG8m融合的突变起始密码子进行体内分析，结果显示缺乏下游重新起始。
Mol Gen Genet. 2000 Jan;262(6):1036-46. doi: 10.1007/pl00008646.
4
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Methods Enzymol. 1996;264:265-78. doi: 10.1016/s0076-6879(96)64026-9.
5
Expression of a recoded nuclear gene inserted into yeast mitochondrial DNA is limited by mRNA-specific translational activation.插入酵母线粒体DNA中的经过重新编码的核基因的表达受到mRNA特异性翻译激活的限制。
Proc Natl Acad Sci U S A. 1996 May 28;93(11):5253-7. doi: 10.1073/pnas.93.11.5253.
6
Patterns of mitochondrial sorting in yeast zygotes.酵母合子中线粒体分选模式。
Mol Biol Cell. 1993 Jan;4(1):21-36. doi: 10.1091/mbc.4.1.21.
7
Suppression of a defect in the 5' untranslated leader of mitochondrial COX3 mRNA by a mutation affecting an mRNA-specific translational activator protein.通过影响一种mRNA特异性翻译激活蛋白的突变抑制线粒体COX3 mRNA 5'非翻译前导区的缺陷。
Mol Cell Biol. 1993 Aug;13(8):4806-13. doi: 10.1128/mcb.13.8.4806-4813.1993.
8
Isolation of the beta-tubulin gene from yeast and demonstration of its essential function in vivo.从酵母中分离β-微管蛋白基因并证明其在体内的重要功能。
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9
Yeast/E. coli shuttle vectors with multiple unique restriction sites.具有多个独特限制性酶切位点的酵母/大肠杆菌穿梭载体。
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10
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