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Study on the construction of recombined plasmid pMG36e-lacc1 and the electroporation of Lactobacillus buchneri

Issue title: Frontiers in Biomedical Engineering and Biotechnology – Proceedings of the 3rd International Conference on Biomedical Engineering and Biotechnology, 25–28 September 2014, Beijing, China

Article type: Research Article

Authors: Xue, Dan | Na, Ri; | Guo, Jiu Feng | Liu, Teng

Affiliations: Key Laboratory of Ion Beam Bioengineering, Inner Mongolia University, Hohhot, China

Note: [] Corresponding author: Ri Na, Key Laboratory of Ion Beam Bioengineering, Inner Mongolia University, Hohhot, China. Tel.:13171079869; Fax: 0471-4993139; E-mail: guojiufeng@imu.edu.cn.

Keywords: Laccase, Lactobacillus buchneri, expression vector system

DOI: 10.3233/BME-141216

Journal: Bio-Medical Materials and Engineering, vol. 24, no. 6, pp. 3855-3861, 2014

Published: 2014
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Abstract

With a goal of obtaining the engineering probiotics that can produce both lactic acid and laccase, Pleurotus eryngii was selected as the test material. First, the laccase gene (Lacc1) was cloned by using RT-PCR (the length of which is 1596bp), and then, the gene was ligated to the food-grade vector pMG36e from Lactobacillus buchneri. The recombinant expression vector pMG36e-Lacc1 was constructed by transforming it into Lactobacillus buchneri via an electroporation method. The recombinant plasmid was constructed successfully as confirmed by gel electrophoresis. Then, the optimum conditions for electroporation were determined. The research revealed that when the electric field intensity is 1.75 kV with an SMRS medium recovery for 1.5 h, the electroporation translation efficiency reaches its maximum level.

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