Use of cyclodextrin and its derivatives for increased transformation efficiency of competent bacterial cells

被引:13
作者
Aachmann, Finn Lillelund [1 ]
Aune, Trond Erik Vee [1 ]
机构
[1] Norwegian Univ Sci & Technol, Dept Biotechnol, NOBIPOL, N-7491 Trondheim, Norway
关键词
Cyclodextrins; DNA; Plasmid; Bacteria; Competent cell; Transformation; ESCHERICHIA-COLI; BETA-CYCLODEXTRIN; CHOLESTEROL DEPLETION; VIRUS TYPE-1; CONSTANT; CARRIER; EFFLUX; DNA;
D O I
10.1007/s00253-009-1907-x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Methodologies for introduction of DNA into cells are essential in molecular genetics and vital for applications such as genetic engineering and gene therapy. The use of cyclodextrins (CyDs) for increased efficiency of introducing DNA into eukaryotic cells (transfection) has been reported, but CyDs' effect on the introduction of DNA into bacterial cells (transformation) is unknown. Here, we have investigated the potential of using CyDs in the transformation of chemically competent in-house, commercially available, and, on non-competent bacterial cells, with plasmid DNA of two different sizes. Possible interactions between CyDs and DNA were studied with nuclear magnetic resonance (NMR) spectroscopy. The presence of CyDs resulted in an up to fourfold increment of the transformation rate for in-house cells, with beta-CyD and derivates giving the strongest effect. For commercial cells and transformation with megaplasmids, a more moderate effect around 1.4-fold was obtained. However, CyDs have little or no effect on DNA uptake by noncompetent cells. Results obtained from NMR spectroscopy show no interactions between CyDs and DNA-like molecules, which indicated that the CyDs' effect is related to the bacterial cell wall.
引用
收藏
页码:589 / 596
页数:8
相关论文
共 38 条
[1]  
[Anonymous], 1998, ESSENTIALS CARBOHYDR
[2]  
[Anonymous], 2012, Molecular Cloning: A Laboratory Manual
[3]   BACTERIAL TOXICITY OF CYCLODEXTRINS - LUMINOUS ESCHERICHIA-COLI AS A MODEL [J].
BAR, R ;
ULITZUR, S .
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, 1994, 41 (05) :574-577
[4]  
BAR R, 1989, APPL MICROBIOL BIOT, V31, P25
[5]   Physicochemical and biological characterization of targeted, nucleic acid-containing nanoparticles [J].
Bartlett, Derek W. ;
Davis, Mark E. .
BIOCONJUGATE CHEMISTRY, 2007, 18 (02) :456-468
[6]  
Chadha R, 2008, J SCI IND RES INDIA, V67, P185
[7]   NONCHROMOSOMAL ANTIBIOTIC RESISTANCE IN BACTERIA - GENETIC TRANSFORMATION OF ESCHERICHIA-COLI BY R-FACTOR DNA [J].
COHEN, SN ;
CHANG, ACY ;
HSU, L .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1972, 69 (08) :2110-&
[8]   CONSTRUCTION OF BIOLOGICALLY FUNCTIONAL BACTERIAL PLASMIDS IN-VITRO [J].
COHEN, SN ;
CHANG, ACY ;
BOYER, HW ;
HELLING, RB .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1973, 70 (11) :3240-3244
[9]   The stability of cyclodextrin complexes in solution [J].
Connors, KA .
CHEMICAL REVIEWS, 1997, 97 (05) :1325-1357
[10]  
Easton C.J., 1999, MODIFIED CYCLODEXTRI