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1.
Nature ; 582(7813): 561-565, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32365353

RESUMO

Reverse genetics has been an indispensable tool to gain insights into viral pathogenesis and vaccine development. The genomes of large RNA viruses, such as those from coronaviruses, are cumbersome to clone and manipulate in Escherichia coli owing to the size and occasional instability of the genome1-3. Therefore, an alternative rapid and robust reverse-genetics platform for RNA viruses would benefit the research community. Here we show the full functionality of a yeast-based synthetic genomics platform to genetically reconstruct diverse RNA viruses, including members of the Coronaviridae, Flaviviridae and Pneumoviridae families. Viral subgenomic fragments were generated using viral isolates, cloned viral DNA, clinical samples or synthetic DNA, and these fragments were then reassembled in one step in Saccharomyces cerevisiae using transformation-associated recombination cloning to maintain the genome as a yeast artificial chromosome. T7 RNA polymerase was then used to generate infectious RNA to rescue viable virus. Using this platform, we were able to engineer and generate chemically synthesized clones of the virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)4, which has caused the recent pandemic of coronavirus disease (COVID-19), in only a week after receipt of the synthetic DNA fragments. The technical advance that we describe here facilitates rapid responses to emerging viruses as it enables the real-time generation and functional characterization of evolving RNA virus variants during an outbreak.


Assuntos
Betacoronavirus/genética , Clonagem Molecular/métodos , Infecções por Coronavirus/virologia , Genoma Viral/genética , Genômica/métodos , Pneumonia Viral/virologia , Genética Reversa/métodos , Biologia Sintética/métodos , Animais , China/epidemiologia , Chlorocebus aethiops , Cromossomos Artificiais de Levedura/metabolismo , Infecções por Coronavirus/epidemiologia , RNA Polimerases Dirigidas por DNA/metabolismo , Evolução Molecular , Humanos , Mutação , Pandemias/estatística & dados numéricos , Pneumonia Viral/epidemiologia , Vírus Sinciciais Respiratórios/genética , Saccharomyces cerevisiae/genética , Células Vero , Proteínas Virais/metabolismo , Zika virus/genética
2.
PLoS One ; 14(8): e0217532, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31412036

RESUMO

Transcriptome analyses show a surprisingly large proportion of the mammalian genome is transcribed; much more than can be accounted for by genes and introns alone. Most of this transcription is non-coding in nature and arises from intergenic regions, often overlapping known protein-coding genes in sense or antisense orientation. The functional relevance of this widespread transcription is unknown. Here we characterize a promoter responsible for initiation of an intergenic transcript located approximately 3.3 kb and 10.7 kb upstream of the adult-specific human ß-globin genes. Mutational analyses in ß-YAC transgenic mice show that alteration of intergenic promoter activity results in ablation of H3K4 di- and tri-methylation and H3 hyperacetylation extending over a 30 kb region immediately downstream of the initiation site, containing the adult δ- and ß-globin genes. This results in dramatically decreased expression of the adult genes through position effect variegation in which the vast majority of definitive erythroid cells harbor inactive adult globin genes. In contrast, expression of the neighboring ε- and γ-globin genes is completely normal in embryonic erythroid cells, indicating a developmentally specific variegation of the adult domain. Our results demonstrate a role for intergenic non-coding RNA transcription in the propagation of histone modifications over chromatin domains and epigenetic control of ß-like globin gene transcription during development.


Assuntos
Cromatina/genética , DNA Intergênico/genética , Regulação da Expressão Gênica no Desenvolvimento , Histonas/química , Regiões Promotoras Genéticas , RNA não Traduzido/genética , Globinas beta/genética , Adulto , Animais , Cromossomos Artificiais de Levedura , Células Eritroides/metabolismo , Humanos , Camundongos , Camundongos Transgênicos , Transcrição Genética
3.
Microb Cell Fact ; 18(1): 52, 2019 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-30857530

RESUMO

BACKGROUND: Strains with increased alkali tolerance have a broad application in industrial, especially for bioremediation, biodegradation, biocontrol and production of bio-based chemicals. A novel synthetic chromosome recombination and modification by LoxP-mediated evolution (SCRaMbLE) system has been introduced in the synthetic yeast genome (Sc 2.0), which enables generation of a yeast library with massive structural variations and potentially drives phenotypic evolution. The structural variations including deletion, inversion and duplication have been detected within synthetic yeast chromosomes. RESULTS: Haploid yeast strains harboring either one (synV) or two (synV and synX) synthetic chromosomes were subjected to SCRaMbLE. Seven of evolved strains with increased alkali tolerance at pH 8.0 were generated through multiple independent SCRaMbLE experiments. Various of structural variations were detected in evolved yeast strains by PCRTag analysis and whole genome sequencing including two complex structural variations. One possessed an inversion of 20,743 base pairs within which YEL060C (PRB1) was deleted simultaneously, while another contained a duplication region of 9091 base pairs in length with a deletion aside. Moreover, a common deletion region with length of 11,448 base pairs was mapped in four of the alkali-tolerant strains. We further validated that the deletion of YER161C (SPT2) within the deleted region could increase alkali tolerance in Saccharomyces cerevisiae. CONCLUSIONS: SCRaMbLE system provides a simple and efficient way to generate evolved yeast strains with enhanced alkali tolerance. Deletion of YER161C (SPT2) mapped by SCRaMbLE can improve alkali tolerance in S. cerevisiae. This study enriches our understanding of alkali tolerance in yeast and provides a standard workflow for the application of SCRaMbLE system to generate various phenotypes that may be interesting for industry and extend understanding of phenotype-genotype relationship.


Assuntos
Álcalis/metabolismo , Cromossomos Artificiais de Levedura/genética , Genoma Fúngico , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Regulação Fúngica da Expressão Gênica , Engenharia Genética , Fenótipo , Recombinação Genética , Proteínas de Saccharomyces cerevisiae/genética , Deleção de Sequência , Biologia Sintética
5.
Nat Commun ; 9(1): 3783, 2018 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-30224715

RESUMO

Structural variations (SVs) exert important functional impacts on biological phenotypic diversity. Here we show a ring synthetic yeast chromosome V (ring_synV) can be used to continuously generate complex genomic variations and improve the production of prodeoxyviolacein (PDV) by applying Synthetic Chromosome Recombination and Modification by LoxP-mediated Evolution (SCRaMbLE) in haploid yeast cells. The SCRaMbLE of ring_synV generates aneuploid yeast strains with increased PDV productivity, and we identify aneuploid chromosome I, III, VI, XII, XIII, and ring_synV. The neochromosome of SCRaMbLEd ring_synV generated more unbalanced forms of variations, including duplication, insertions, and balanced forms of translocations and inversions than its linear form. Furthermore, of the 29 novel SVs detected, 11 prompted the PDV biosynthesis; and the deletion of uncharacterized gene YER182W is related to the improvement of the PDV. Overall, the SCRaMbLEing ring_synV embraces the evolution of the genome by modifying the chromosome number, structure, and organization, identifying targets for phenotypic comprehension.


Assuntos
Cromossomos Artificiais de Levedura , Engenharia Genética/métodos , Saccharomyces cerevisiae/genética , Aneuploidia , Deleção de Genes , Variação Genética , Genoma Fúngico , Genótipo , Haploidia , Indóis/metabolismo , Microrganismos Geneticamente Modificados , Fenótipo , Reação em Cadeia da Polimerase/métodos , Saccharomyces cerevisiae/metabolismo
6.
Nature ; 560(7718): 331-335, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-30069045

RESUMO

Eukaryotic genomes are generally organized in multiple chromosomes. Here we have created a functional single-chromosome yeast from a Saccharomyces cerevisiae haploid cell containing sixteen linear chromosomes, by successive end-to-end chromosome fusions and centromere deletions. The fusion of sixteen native linear chromosomes into a single chromosome results in marked changes to the global three-dimensional structure of the chromosome due to the loss of all centromere-associated inter-chromosomal interactions, most telomere-associated inter-chromosomal interactions and 67.4% of intra-chromosomal interactions. However, the single-chromosome and wild-type yeast cells have nearly identical transcriptome and similar phenome profiles. The giant single chromosome can support cell life, although this strain shows reduced growth across environments, competitiveness, gamete production and viability. This synthetic biology study demonstrates an approach to exploration of eukaryote evolution with respect to chromosome structure and function.


Assuntos
Cromossomos Artificiais de Levedura/genética , Engenharia Genética/métodos , Aptidão Genética/genética , Saccharomyces cerevisiae/citologia , Saccharomyces cerevisiae/genética , Biologia Sintética/métodos , Fusão Gênica Artificial/métodos , Centrômero/genética , Evolução Molecular , Meiose , Viabilidade Microbiana/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento , Esporos Fúngicos/genética , Telômero/genética , Transcriptoma
7.
Cold Spring Harb Protoc ; 2018(8)2018 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-30068584

RESUMO

This protocol describes a method to purify yeast artificial chromosome (YAC) DNA for microinjection. YAC DNA solutions cannot be concentrated by standard DNA precipitation methods and resuspended into smaller volumes. Attempts to precipitate YAC DNA solutions will, unavoidably, break and destroy the DNA molecules. Therefore YAC DNA must always be kept in a solution that maintains the integrity of the YAC DNA molecules. In the method described here, YAC DNA is extracted from pulsed-field gel electrophoresis (PFGE) agarose plugs by using two gel electrophoresis steps. The first gel electrophoresis step is the PFGE itself. The region of agarose containing the YAC DNA of interest is excised and run on a standard electrophoresis gel setup at a 90° angle to the PFGE run. The YAC DNA migrates out of the agarose slice and enters into a thicker low-melting agarose gel, thereby promoting compaction of YAC DNA molecules. The aim of this method is to convert a slice of agarose into a cube of agarose of smaller volume. The volume of this cube will determine the final concentration of the YAC DNA precipitation.


Assuntos
Cromossomos Artificiais de Levedura , DNA Fúngico/isolamento & purificação , Eletroforese em Gel Bidimensional/métodos , Microinjeções , Eletroforese em Gel de Ágar/métodos , Eletroforese em Gel de Campo Pulsado/métodos
8.
Cold Spring Harb Protoc ; 2018(8)2018 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-30068585

RESUMO

This protocol describes a method for concentrating yeast artificial chromosome (YAC) DNA preparations following separation of chromosomes by pulsed-field gel electrophoresis (PFGE). The agarose slice containing the YAC DNA of interest is melted and digested. The resulting YAC DNA-containing solution is concentrated using ultrafiltration units with a 30,000-mwt cutoff. The DNA is retained in the upper container, where it concentrates sixfold to sevenfold. Ideally, YAC DNA preparations at 10-60 ng/µL can be obtained. However, great care must be taken to recover the YAC DNA molecules that sometimes irreversibly attach to the filter membrane. This procedure can also be used to concentrate BAC DNA from PFGE gel slices of BAC DNA-containing agarose exactly as described for YACs.


Assuntos
Cromossomos Artificiais de Levedura , DNA Fúngico/isolamento & purificação , Eletroforese em Gel de Campo Pulsado/métodos , Ultrafiltração/métodos , Cromossomos Artificiais Bacterianos , Sefarose
9.
Cold Spring Harb Protoc ; 2018(8)2018 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-30068586

RESUMO

The microinjection of DNA directly into the pronuclei of fertilized zygotes is the most extensively used method of gene transfer in the mouse. The injection of very large pieces of DNA, including bacterial artificial chromosomes (BACs) and yeast artificial chromosomes (YACs), has become increasingly popular because their size normally accommodates all of the regulatory elements that are needed for a given expression domain to function adequately in an ectopic genomic location. This protocol describes how to prepare large-scale yeast agarose plugs to isolate YAC DNA. The quality of the YAC DNA preparation is the most important consideration in optimizing transgenic mouse production.


Assuntos
Cromossomos Artificiais de Levedura , DNA Fúngico/isolamento & purificação , Sefarose/síntese química , Animais , Camundongos , Camundongos Transgênicos , Microinjeções
10.
J Microbiol Biotechnol ; 28(5): 821-825, 2018 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-29551023

RESUMO

A copy number amplification system for yeast artificial chromosomes (YACs) was combined with simultaneous overexpression of genes integrated into a YAC. The chromosome VII (1,105 kb) was successfully split to 887 kb, 44 kb containing the element for copy number amplification, and a 184-kb split-YAC. The 44-kb split-mini YAC was amplified a maximum of 9-fold, and the activity of the reporter enzymes integrated into the split-mini YAC increased about 5-7-fold. These results demonstrate that the mini-YAC containing a targeted chromosome region can be readily amplified, and the specific genes in the mini-YAC could be overexpressed by increasing the copy number.


Assuntos
Cromossomos Artificiais de Levedura/genética , Dosagem de Genes/genética , Saccharomyces cerevisiae/genética , Cromossomos Fúngicos/genética , Eletroforese em Gel de Campo Pulsado , Reação em Cadeia da Polimerase
11.
Methods Mol Biol ; 1698: 37-65, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29076083

RESUMO

Animal models of erythropoiesis have been, and will continue to be, important tools for understanding molecular mechanisms underlying the development of this cell lineage and the pathophysiology associated with various human erythropoietic diseases. In this regard, the mouse is probably the most valuable animal model available to investigators. The physiology and short gestational period of mice make them ideal for studying developmental processes and modeling human diseases. These attributes, coupled with cutting-edge genetic tools such as transgenesis, gene knockouts, conditional gene knockouts, and genome editing, provide a significant resource to the research community to test a plethora of hypotheses. This review summarizes the mouse models available for studying a wide variety of erythroid-related questions, as well as the properties inherent in each one.


Assuntos
Eritropoese , Hemoglobinopatias/genética , Animais , Cromossomos Artificiais Bacterianos , Cromossomos Artificiais de Levedura , Modelos Animais de Doenças , Regulação da Expressão Gênica no Desenvolvimento , Genes Reporter , Hemoglobinopatias/metabolismo , Hemoglobinopatias/patologia , Hemoglobinas/genética , Camundongos , Camundongos Knockout , Camundongos Transgênicos
12.
Methods Mol Biol ; 1672: 403-419, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29043639

RESUMO

Trinucleotide repeats are common in the human genome and can undergo changes in repeat length. Expanded CAG repeats have been linked to over 14 human diseases and are considered hotspots for breakage and genomic rearrangement. Here, we describe two Saccharomyces cerevisiae based assays that evaluate the rate of chromosome breakage that occurs within a repeat tract (fragility), and a PCR-based assay to evaluate tract length changes (instability). The first fragility assay utilizes end-loss and subsequent telomere addition as the main mode of repair of a yeast artificial chromosome (YAC). The second fragility assay relies on the fact that a chromosomal break stimulates recombination-mediated repair. In addition to understanding the role of fragility at repetitive DNA sequences, both assays can be modified to evaluate instability of a CAG repeat using a PCR-based assay. All three assays have been essential in understanding the genetic mechanisms that cause chromosome breaks and tract-length changes at unstable repeats.


Assuntos
Instabilidade Genômica , Sequências Repetitivas de Ácido Nucleico , Saccharomyces cerevisiae/genética , Quebra Cromossômica , Cromossomos Artificiais de Levedura , Recombinação Genética , Expansão das Repetições de Trinucleotídeos , Repetições de Trinucleotídeos
13.
Yi Chuan ; 39(10): 865-876, 2017 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-29070482

RESUMO

With the rapid growth and development of synthetic biology, research in the genomics is advancing from genome sequencing to genome synthesis. In 2009, Professor Jef D. Boeke proposed the Synthetic Yeast Genome Project (Sc2.0), which aims to synthesize the world's first eukaryotic genome. With the efforts of scientists from the United States, China, Britain, France, Australia, Singapore and other countries, a third of the Saccharomyces cerevisiae chromosomes has now been synthesized. In the perspectives of synthetic genomics, we here review the recent progress in the Sc2.0 project, including discussion on the right arm of chromosome 9, and chromosomes 2, 5, 6, 10, 12, in terms of their designs and synthetic strategy as well as the biological significance, thereby providing a reference for further research in synthetic genomics.


Assuntos
Cromossomos Artificiais de Levedura , Saccharomyces cerevisiae/genética , Genoma Fúngico , Biologia Sintética
15.
Science ; 355(6329)2017 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-28280149

RESUMO

We designed and synthesized a 976,067-base pair linear chromosome, synXII, based on native chromosome XII in Saccharomyces cerevisiae SynXII was assembled using a two-step method, specified by successive megachunk integration and meiotic recombination-mediated assembly, producing a functional chromosome in S. cerevisiae. Minor growth defect "bugs" detected in synXII, caused by deletion of tRNA genes, were rescued by introducing an ectopic copy of a single tRNA gene. The ribosomal gene cluster (rDNA) on synXII was left intact during the assembly process and subsequently replaced by a modified rDNA unit used to regenerate rDNA at three distinct chromosomal locations. The signature sequences within rDNA, which can be used to determine species identity, were swapped to generate a Saccharomyces synXII strain that would be identified as Saccharomyces bayanus by standard DNA barcoding procedures.


Assuntos
Cromossomos Artificiais de Levedura/química , DNA Ribossômico/genética , Engenharia Genética/métodos , Genoma Fúngico , Saccharomyces cerevisiae/genética , Biologia Sintética/métodos , Núcleo Celular/genética , Núcleo Celular/ultraestrutura , Cromossomos Artificiais de Levedura/genética , Cromossomos Artificiais de Levedura/ultraestrutura , Saccharomyces cerevisiae/ultraestrutura , Transcriptoma
16.
Science ; 355(6329)2017 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-28280150

RESUMO

Although the design of the synthetic yeast genome Sc2.0 is highly conservative with respect to gene content, the deletion of several classes of repeated sequences and the introduction of thousands of designer changes may affect genome organization and potentially alter cellular functions. We report here the Hi-C-determined three-dimensional (3D) conformations of Sc2.0 chromosomes. The absence of repeats leads to a smoother contact pattern and more precisely tractable chromosome conformations, and the large-scale genomic organization is globally unaffected by the presence of synthetic chromosome(s). Two exceptions are synIII, which lacks the silent mating-type cassettes, and synXII, specifically when the ribosomal DNA is moved to another chromosome. We also exploit the contact maps to detect rearrangements induced in SCRaMbLE (synthetic chromosome rearrangement and modification by loxP-mediated evolution) strains.


Assuntos
Cromossomos Artificiais de Levedura/ultraestrutura , Genoma Fúngico , Saccharomyces cerevisiae/genética , Biologia Sintética , Núcleo Celular/genética , Núcleo Celular/ultraestrutura , Centrômero/ultraestrutura , Cromossomos Artificiais de Levedura/química , Cromossomos Artificiais de Levedura/genética , DNA Ribossômico/genética , Conformação de Ácido Nucleico , Sequências Repetitivas de Ácido Nucleico/genética , Deleção de Sequência , Telômero/ultraestrutura
17.
Science ; 355(6329)2017 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-28280151

RESUMO

Perfect matching of an assembled physical sequence to a specified designed sequence is crucial to verify design principles in genome synthesis. We designed and de novo synthesized 536,024-base pair chromosome synV in the "Build-A-Genome China" course. We corrected an initial isolate of synV to perfectly match the designed sequence using integrative cotransformation and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated editing in 22 steps; synV strains exhibit high fitness under a variety of culture conditions, compared with that of wild-type V strains. A ring synV derivative was constructed, which is fully functional in Saccharomyces cerevisiae under all conditions tested and exhibits lower spore viability during meiosis. Ring synV chromosome can extends Sc2.0 design principles and provides a model with which to study genomic rearrangement, ring chromosome evolution, and human ring chromosome disorders.


Assuntos
Cromossomos Artificiais de Levedura/química , Genoma Fúngico , Saccharomyces cerevisiae/genética , Biologia Sintética/métodos , Proteínas de Bactérias , Proteína 9 Associada à CRISPR , Cromossomos Artificiais de Levedura/genética , Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas , Endonucleases , Edição de Genes , Rearranjo Gênico , Meiose , Modelos Genéticos , Saccharomyces cerevisiae/citologia , Transformação Genética
18.
Science ; 355(6329)2017 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-28280152

RESUMO

Debugging a genome sequence is imperative for successfully building a synthetic genome. As part of the effort to build a designer eukaryotic genome, yeast synthetic chromosome X (synX), designed as 707,459 base pairs, was synthesized chemically. SynX exhibited good fitness under a wide variety of conditions. A highly efficient mapping strategy called pooled PCRTag mapping (PoPM), which can be generalized to any watermarked synthetic chromosome, was developed to identify genetic alterations that affect cell fitness ("bugs"). A series of bugs were corrected that included a large region bearing complex amplifications, a growth defect mapping to a recoded sequence in FIP1, and a loxPsym site affecting promoter function of ATP2 PoPM is a powerful tool for synthetic yeast genome debugging and an efficient strategy for phenotype-genotype mapping.


Assuntos
Cromossomos Artificiais de Levedura/química , Cromossomos Artificiais de Levedura/genética , Genoma Fúngico , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Mapeamento Físico do Cromossomo/métodos , Saccharomyces cerevisiae/genética , Sequência de Bases , Duplicação Gênica , Aptidão Genética , Biologia Sintética
19.
Science ; 355(6329)2017 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-28280153

RESUMO

Here, we report the successful design, construction, and characterization of a 770-kilobase synthetic yeast chromosome II (synII). Our study incorporates characterization at multiple levels-including phenomics, transcriptomics, proteomics, chromosome segregation, and replication analysis-to provide a thorough and comprehensive analysis of a synthetic chromosome. Our Trans-Omics analyses reveal a modest but potentially relevant pervasive up-regulation of translational machinery observed in synII, mainly caused by the deletion of 13 transfer RNAs. By both complementation assays and SCRaMbLE (synthetic chromosome rearrangement and modification by loxP-mediated evolution), we targeted and debugged the origin of a growth defect at 37°C in glycerol medium, which is related to misregulation of the high-osmolarity glycerol response. Despite the subtle differences, the synII strain shows highly consistent biological processes comparable to the native strain.


Assuntos
Cromossomos Artificiais de Levedura/fisiologia , Genoma Fúngico , Saccharomyces cerevisiae/genética , Segregação de Cromossomos , Cromossomos Artificiais de Levedura/química , Cromossomos Artificiais de Levedura/genética , Meios de Cultura/química , Replicação do DNA , Glicerol , Proteômica , Saccharomyces cerevisiae/crescimento & desenvolvimento , Análise de Sequência de DNA , Biologia Sintética , Transcriptoma
20.
Science ; 355(6329)2017 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-28280154

RESUMO

We describe design, rapid assembly, and characterization of synthetic yeast Sc2.0 chromosome VI (synVI). A mitochondrial defect in the synVI strain mapped to synonymous coding changes within PRE4 (YFR050C), encoding an essential proteasome subunit; Sc2.0 coding changes reduced Pre4 protein accumulation by half. Completing Sc2.0 specifies consolidation of 16 synthetic chromosomes into a single strain. We investigated phenotypic, transcriptional, and proteomewide consequences of Sc2.0 chromosome consolidation in poly-synthetic strains. Another "bug" was discovered through proteomic analysis, associated with alteration of the HIS2 transcription start due to transfer RNA deletion and loxPsym site insertion. Despite extensive genetic alterations across 6% of the genome, no major global changes were detected in the poly-synthetic strain "omics" analyses. This work sets the stage for completion of a designer, synthetic eukaryotic genome.


Assuntos
Cromossomos Artificiais de Levedura/química , Cromossomos Artificiais de Levedura/genética , Saccharomyces cerevisiae/genética , Biologia Sintética/métodos , Células Artificiais/metabolismo , Mapeamento Físico do Cromossomo , Complexo de Endopeptidases do Proteassoma/genética , Proteômica , Saccharomyces cerevisiae/crescimento & desenvolvimento , Proteínas de Saccharomyces cerevisiae/genética
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