在大肠杆菌基因组中用2'-deoxycytidine类似物取代2'-deoxycytidine
Angad P Mehta1, Han Li1, Sean A Reed1
1The Scripps Research Institute , 10550 North Torrey Pines Road, La Jolla, California 92037, United States.
Journal of the American Chemical Society
|November 3, 2016
概括
研究人员对大肠杆菌进行了基因工程,将63%的二氧化 (dC) 替换为5-甲基二氧化 (5hmC),并在表观遗传调节研究中获得了20%的基因组5-甲基二氧化 (5-gmC).
科学领域:
- 合成生物学
- 表观遗传学
- 微生物遗传学
背景情况:
- 在DNA中修改的核酸对表观遗传调节和限制系统至关重要.
- 在之前的研究中,在大肠杆菌中用5 extquotesingle-hydroxymethyl-2 extquotesingle-deoxyuridine (5hmU) 取代了75%的提米丁 (T).
研究的目的:
- 设计大肠杆菌中的胺核酸生物合成途径.
- 为了研究用5-基-2-基-基 (5hmC) 替换2基-基-基.
- 进一步将5hmC改为5-glucosyl-5-hydroxymethyl-2 extquotesingle-deoxycytidine (5-gmC),并评估其基因组和等离子体DNA的整合.
主要方法:
- 利用 T4 菌体基因来设计胺核酸生物合成途径.
- 改造了葡萄糖代谢途径以促进核酸变异.
- 在基因组和等离子体DNA中使用分子生物学技术量化核酸替代水平.
主要成果:
- 在大肠杆菌基因组中,大约63%的dC被5hmC取代.
- 在等离子体DNA中,大约有71%的dC被5hmC取代.
- 在基因组DNA中产生20%的5-gmC,在等离子体DNA中产生45%的5-gmC.
结论:
- 证明了大肠杆菌的成功工程,用于高水平的修饰细胞因子衍生物.
- 建立了一个生成5gmC修饰DNA的系统,为研究其生物功能铺平了道路.
- 提供了解细菌基因组广泛核酸修饰的后果的基础.
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