Effects of Sr2+ on the preparation of Escherchia coli DH5 alpha competent cells and plasmid transformation

doi:10.7717/peerj.9480

	Effects of Sr2+ on the preparation of Escherchia coli DH5 alpha competent cells and plasmid transformation
	Wang, Yonggang 1,2; Wang, Xinjian 2; Yu, Linmiao 2; Tian, Yuan 3; Li, Shaowei 3; Leng, Feifan 2; Ma, Jianzhong 2; Chen, Jixiang 1,4
刊名	PEERJ
	2020-08-13
卷号	8
关键词	Escherchia coli Response surface methodology Competence Transformation mechanism
ISSN号	2167-8359
DOI	10.7717/peerj.9480
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研究方向	Science & Technology - Other Topics
语种	英语
出版者	PEERJ INC
WOS记录号	WOS:000558958100003
内容类型	期刊论文
源URL	[http://ir.lut.edu.cn/handle/2XXMBERH/155123]
专题	生命科学与工程学院石油化工学院
作者单位	1.Lanzhou Univ Technol, Sch Energy & Power Engn, Lanzhou, Gansu, Peoples R China; 2.Lanzhou Univ Technol, Sch Life Sci & Engn, Lanzhou, Gansu, Peoples R China; 3.Chinese Acad Sci, Inst Geog Sci & Nat Resources Res, Key Lab Ecosyst Network Observat & Modelling, Lhasa Natl Ecol Res Stn, Beijing, 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.
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.
MLA	Wang, Yonggang,et al."Effects of Sr2+ on the preparation of Escherchia coli DH5 alpha competent cells and plasmid transformation".PEERJ 8(2020).