title: Study on the construction of recombined plasmid pMG36e-lacc1 and the electroporation of Lactobacillus buchneri.
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 1596 bp), 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.
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Inner Mongolia University, Hohhot, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Xue
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Inner Mongolia University, Hohhot, China.
authors.forename: Ri
authors.initials: R
authors.lastname: Na
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Inner Mongolia University, Hohhot, China.
authors.forename: Jiu Feng
authors.initials: JF
authors.lastname: Guo
authors.affiliation: Key Laboratory of Ion Beam Bioengineering, Inner Mongolia University, Hohhot, China.
authors.forename: Teng
authors.initials: T
authors.lastname: Liu
mesh_terms: D003001:Cloning, Molecular / Q000379:methods; D015195:Drug Design; D018274:Electroporation / Q000379:methods*; D042845:Laccase / Q000235:genetics*; D007778:Lactobacillus / Q000235:genetics*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics*; D020076:Pleurotus / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics*; D013329:Structure-Activity Relationship; D014162:Transfection / Q000379:methods*
chemical_list: D042845:Laccase
keywords: Laccase; Lactobacillus buchneri; expression vector system