Effects of Sr2+ on the preparation of Escherchia coli DH5 alpha competent cells and plasmid transformation
文献类型：期刊论文
| 作者 | Wang, Yonggang2,3; Wang, Xinjian3; Yu, Linmiao3; Tian, Yuan1; Li, Shaowei1; Leng, Feifan3; Ma, Jianzhong3; Chen, Jixiang2,4 |
| 刊名 | PEERJ
 |
| 出版日期 | 2020-08-13 |
| 卷号 | 8页码:20 |
| 关键词 | Escherchia coli Response surface methodology Competence Transformation mechanism |
| ISSN号 | 2167-8359 |
| DOI | 10.7717/peerj.9480 |
| 通讯作者 | Wang, Yonggang(wangyg@lut.cn) ; Chen, Jixiang(chenjixiang@lut.cn) |
| 英文摘要 | Bacterial gene transformation used with Escherichia coli as a desired microorganism is one of the important techniques in genetic engineering. In this study, the preparation of E. coli DH5 alpha competent cells treated with SrCl2 and transformation by heat-shock with pUC19 plasmid was optimized by Response Surface Methodology (RSM). Other five E. coli strains including BL21 (DE3), HB-101, JM109, TOP10 and TG1, three different sizes plasmids (pUC19, pET32a, pPIC9k) were used to verify the protocol, respectively. The transformation mechanism was explored by scanning electron microscope combined with energy dispersive spectrometer (SEM-EDS), atomic absorption spectroscopy (AAS) and Fourier-transform infrared spectroscopy (FT-IR). An equation of regression model was obtained, and the ideal parameters were Sr(2+ )ions of 90 mM, heat-shock time of 90 s and 9 ng of plasmid. Under this conditions, the transformation efficiency could almost reach to 10(6) CFU/mu g DNA. A small change of the cell surface structure has been observed between E. coli DH5 alpha strain and competent cells by abovementioned spectrum technologies, which implied that a strict regulation mechanism involved in the formation of competent cells and transformation of plasmids. An equation of regression model for the competent cells preparation and plasmid transformation could be applied in gene cloning technology |
| WOS关键词 | HIGH-EFFICIENCY TRANSFORMATION ; ESCHERICHIA-COLI ; ARTIFICIAL TRANSFORMATION ; CALCIUM-CHLORIDE ; DNA ; OPTIMIZATION ; MECHANISM ; STORAGE ; RANGE ; TOOL |
| 资助项目 | National Natural Science Foundation of China[31760028] ; Fundamental Research Funds for Key Laboratory of Drug Screening and Deep Processing for Traditional Chinese and Tibetan Medicine of Gansu Province[KZZY20180605] ; Youth talent support program of Lanzhou University and Technology[2018] |
| WOS研究方向 | Science & Technology - Other Topics |
| 语种 | 英语 |
| WOS记录号 | WOS:000558958100003 |
| 出版者 | PEERJ INC |
| 资助机构 | National Natural Science Foundation of China ; Fundamental Research Funds for Key Laboratory of Drug Screening and Deep Processing for Traditional Chinese and Tibetan Medicine of Gansu Province ; Youth talent support program of Lanzhou University and Technology |
| 源URL | [http://ir.igsnrr.ac.cn/handle/311030/158080]  |
| 专题 | 中国科学院地理科学与资源研究所 |
| 通讯作者 | Wang, Yonggang; Chen, Jixiang |
| 作者单位 | 1.Chinese Acad Sci, Inst Geog Sci & Nat Resources Res, Key Lab Ecosyst Network Observat & Modelling, Lhasa Natl Ecol Res Stn, Beijing, Peoples R China 2.Lanzhou Univ Technol, Sch Energy & Power Engn, Lanzhou, Gansu, Peoples R China 3.Lanzhou Univ Technol, Sch Life Sci & Engn, Lanzhou, Gansu, Peoples R China 4.Lanzhou Univ Technol, Sch Petrochem Engn, Lanzhou, Gansu, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wang, Yonggang,Wang, Xinjian,Yu, Linmiao,et al. Effects of Sr2+ on the preparation of Escherchia coli DH5 alpha competent cells and plasmid transformation[J]. PEERJ,2020,8:20. |
| APA | Wang, Yonggang.,Wang, Xinjian.,Yu, Linmiao.,Tian, Yuan.,Li, Shaowei.,...&Chen, Jixiang.(2020).Effects of Sr2+ on the preparation of Escherchia coli DH5 alpha competent cells and plasmid transformation.PEERJ,8,20. |
| MLA | Wang, Yonggang,et al."Effects of Sr2+ on the preparation of Escherchia coli DH5 alpha competent cells and plasmid transformation".PEERJ 8(2020):20. |
入库方式： OAI收割
来源：地理科学与资源研究所
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