Optimization and mechanism exploration for Escherichia coli transformed with plasmid pUC19 by the combination with ultrasound treatment and chemical method.
Optimization and mechanism exploration for Escherichia coli transformed with plasmid pUC19 by the combination with ultrasound treatment and chemical method.
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超声处理与化学法相结合的pUC19质粒转化大肠杆菌的优化及机制探讨
DOI：
10.1016/j.ultsonch.2021.105552
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发表时间：
2021-06
影响因子：
8.4
通讯作者：
Ma J
中科院分区：
文献类型：
--
作者：
Wang Y;Sun S;Yu L;Hu S;Fan W;Leng F;Ma J
优化了超声介导的转移方法。E. coli DH 5 α为受体，pUC 19为外源DNA。急诊coli DH 5 α细胞膜透性发生变化，允许外源DNA进入细胞质。生物转化作为分子克隆的基础技术，已成功地应用于生物医药和食品加工等领域。本研究建立了外源DNA转化E.大肠杆菌细胞。在响应面法优化的最佳转化条件（35 °C，40 W，25 s，OD 600 = 0.4-0.6）下，转化效率可达1.006 × 107 CFU/μg DNA。超声处理前后细胞膜通透性、大分子物质和细胞结构分析结果表明，较低（40 W）超声和较短超声时间（25 s）对宿主细胞的损伤是可逆的，在此条件下对转化效率和细胞存活率无明显影响。总之，细胞膜和细胞壁的适当变化是宿主细胞摄取外源DNA的基本条件，而外源DNA能否在细胞内复制和表达则取决于宿主细胞的活力。
The transfer method was optimized in ultrasound-mediated. E. coli DH5α as the receptor, pUC19 as exogenous DNA. The E. coli DH5α membrane permeability changed allowing exogenous DNA to entered the cytoplasm. As a basic technique of molecular cloning, bio-transformation has been successfully used in the fields of biomedicine and food processing. In this study, we established a transformation system of exogenous DNA into E. coli cells mediated by ultrasound. Under the optimal conditions (i.e. 35 °C, 40 W, 25 s, OD600 = 0.4–0.6) optimized by RSM, the transformation efficiency reached at 1.006 × 107 CFU/μg DNA. The results of membrane permeability, macromolecular substance and cell structure analysis before and after ultrasound treatment showed that the damage of host cells induced by lower (40 W) ultrasound and shorter ultrasound time (25 s) was reversible, and the transformation efficiency and cell survival rate were not significantly affected under this condition. In brief, proper changes in cell membrane and cell wall were the basic conditions for host cells to uptake exogenous DNA, while, whether exogenous DNA could be replicated and expressed in cells depends on the viability of host cells.
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影响因子：
0.7
作者：
Rajagopal, Kammara;Singh, Praveen Kumar;Kumar, Rajesh;Siddiqui, Kaneez Fatima
通讯作者：
Siddiqui, Kaneez Fatima
影响因子：
3.6
作者：
Macfadyen, LP;Chen, D;Redfield, RJ
通讯作者：
Redfield, RJ
影响因子：
3
作者：
Guo, Rongxin;Zheng, Xusheng;Liu, Yanping
通讯作者：
Liu, Yanping
DOI：
10.1073/pnas.86.7.2172
发表时间：
1989-04-01
影响因子：
11.1
作者：
CHUNG, CT;NIEMELA, SL;MILLER, RH
通讯作者：
MILLER, RH
影响因子：
4
作者：
Poyraz, Cagla;Kucukyildiz, Gurkan;Sahin, Selin
通讯作者：
Sahin, Selin