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Method for increasing genetic transformation of improved Shewanella oneidensis MR-1

A technology of genetic transformation and nano-titanium dioxide, which is applied to other methods of inserting foreign genetic materials, recombinant DNA technology, etc., can solve the problems of low efficiency of conjugation transformation methods, and achieve the effects of improving the efficiency of conjugation transformation, simple operation and low cost.

Inactive Publication Date: 2013-10-23
JIANGSU UNIV
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Problems solved by technology

[0004] The purpose of the present invention is to address the traditional Shewanella Due to the low efficiency of the conjugative transformation method of bacteria, an improved method for efficient conjugative transformation was established by adding nano-titanium dioxide as a medium in the transformation system

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  • Method for increasing genetic transformation of improved Shewanella oneidensis MR-1

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Embodiment Construction

[0024] Donor bacteria of the present invention E. coli WM3064 is a commonly used bacterial strain in molecular biology, donated by Dr. Wu Chao of University of Science and Technology of China, and the expression plasmid pBBR1MCS-5 with Gm resistance carried by it (the expression plasmid was purchased from China Plasmid Vector Strain Cell Line Gene Collection Center) was provided by this Laboratory self-transformation; recipient bacteria S. oneidensis MR-1 was donated by Professor Nelson of the University of California, USA.

[0025] 1. (1) The expression plasmid pBBR1MCS-5 of Gm resistance was transformed into Escherichia coli:

[0026] (1) Take 100 μl of WM3064 competent cells (prepared in advance using the Super Competent Cell Preparation Kit produced by Shanghai Sangon Bioengineering Co., Ltd.) and thaw on an ice bath;

[0027] (2) Add 5 μl of pure pBBR1MCS-5 plasmid, mix gently by pipetting, and place in ice bath for 30 min.

[0028] (3) Put the bacteria liquid into ...

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Abstract

The invention relates to a method for increasing genetic transformation of improved Shewanella oneidensis MR-1 and belongs to the field of mutation or genetic engineering. The method comprises the following steps: adding nanometer titanium dioxide grains into a solution formed by mixing a donor bacterium Escherichiacoli WM3064 and a recipient bacterium S.oneidensis MR-1 of a foreign gene in an equal proportion way, and then statically culturing. According to the method, the foreign gene is effectively boosted to transfer from the donor bacterium to the recipient bacterium and the genetic transformation efficiency of the foreign gene to the S.oneidensis MR-1 is obviously increased. The invention also provides a test basis to the new application of a nanometer material in the field of genetic engineering. The method has a potential practical use value and meets the demand of the national strategy of sustainable development.

Description

technical field [0001] The invention relates to a method of improving the anti-genetic effect of exogenous genes by using nano-titanium dioxide as a medium. S. oneidensis The invention relates to an efficient improvement method for genetic transformation of MR-1, which belongs to the field of mutation or genetic engineering. Background technique [0002] Shewanella (Shewanella) is an important environmental microorganism that is widely distributed in nature, especially in aquatic environments. Shewanella Not only is it a multifunctional metal-reducing bacterium, it uses metal oxides in the environment as electron acceptors, and can also use many pollutants in the environment for anaerobic respiration, including inorganic oxides such as nitrite and sulfite, and technetium, Neptunium, plutonium, Cr(VI), U(VI) and vanadate and other radioactive or heavy metal compounds, etc. Studies in recent years have found that, Shewanella Bacteria can also use artificial dyes as elec...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/87
Inventor 肖翔吴勇民曹丹鸣杜道林
Owner JIANGSU UNIV
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