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学科主题: 临床医学
题名:
Whole-Genome Synthesis and Characterization of Viable S13-Like Bacteriophages
作者: Liu, Yuchen1,2; Han, Yonghua1; Huang, Weiren1,4; Duan, Yonggang1; Mou, Lisha1; Jiang, Zhimao1; Fa, Pingping1; Xie, Jun1; Diao, Ruiying1; Chen, Yuanbin1; Ye, Yiwang1; Yang, Ruilin1; Chen, Jing1; Sun, Xiaojuan1; Li, Zesong1; Tang, Aifa1; Gui, Yaoting1; Cai, Zhiming1,3,4
刊名: PLOS ONE
发表日期: 2012-07-18
DOI: 10.1371/journal.pone.0041124
卷: 7, 期:7
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Multidisciplinary Sciences
研究领域[WOS]: Science & Technology - Other Topics
关键词[WOS]: FUNCTIONAL-RELATIONSHIP ; NUCLEOTIDE-SEQUENCE ; EVOLUTION ; PHI-X174 ; PHIX174 ; BIOLOGY ; PHAGES ; DNA ; OLIGONUCLEOTIDES ; GENERATION
英文摘要:

Background: Unprecedented progresses in high-throughput DNA sequencing and de novo gene synthesis technologies have allowed us to create living organisms in the absence of natural template.

Methodology/Principal Findings: The sequence of wild-type S13 phage genome was downloaded from GenBank. Two synonymous mutations were introduced into wt-S13 genome to generate m1-S13 genome. Another mutant, m2-S13 genome, was obtained by engineering two nonsynonymous mutations in the capsid protein coding region of wt-S13 genome. A chimeric phage genome was designed by replacing the F capsid protein open reading frame (ORF) from phage S13 with the F capsid protein ORF from phage G4. The whole genomes of all four phages were assembled from a series of chemically synthesized short overlapping oligonucleotides. The linear synthesized genomes were circularized and electroporated into E. coli C, the standard laboratory host of S13 phage. All four phages were recovered and plaques were visualized. The results of sequencing showed the accuracy of these synthetic genomes. The synthetic phages were capable of lysing their bacterial host and tolerating general environmental conditions. While no phenotypic differences among the variant strains were observed when grown in LB medium with CaCl2, the S13/G4 chimera was found to be much more sensitive to the absence of calcium and to have a lower adsorption rate under calcium free condition.

Conclusions/Significance: The bacteriophage S13 and its variants can be chemically synthesized. The major capsid gene of phage G4 is functional in the phage S13 life cycle. These results support an evolutional hypothesis which has been proposed that a homologous recombination event involving gene F of quite divergent ancestral lineages should be included in the history of the microvirid family.

语种: 英语
所属项目编号: 81100460 ; 20100480776 ; CXB200903090055A ; 200800010106
项目资助者: National Natural Science Foundation of China ; Postdoctoral Science Foundation of China ; Promotion Program for Shenzhen Key Laboratory, Shenzhen, China ; Ph.D. Programs Foundation of Ministry of Education of China
WOS记录号: WOS:000306548900076
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/62189
Appears in Collections:北京大学深圳医院_期刊论文

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作者单位: 1.Shantou Univ, Coll Med, Shantou, Peoples R China
2.Shenzhen Univ, Inst Urogenital Dis, Shenzhen, Peoples R China
3.Peking Univ, Shenzhen Hosp, Shenzhen PKUHKUST Med Ctr,Inst Urol, Guangdong & Shenzhen Key Lab Male Reprod Med & Ge, Shenzhen, Peoples R China
4.Shenzhen Second Peoples Hosp, Shenzhen, Peoples R China

Recommended Citation:
Liu, Yuchen,Han, Yonghua,Huang, Weiren,et al. Whole-Genome Synthesis and Characterization of Viable S13-Like Bacteriophages[J]. PLOS ONE,2012,7(7).
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