A comparison of methods for extracting plasmids from a difficult to lyse bacterium: Lactobacillus casei.
There are few practical protocols to extract efficient plasmid DNA from the difficult-to-lyse bacterium, Lactobacillus casei. This is related to production of a large amount of exopolysaccharide coat and its special physiological characteristics. In this study, we optimized a protocol to extract efficient plasmid DNA from a recombinant L. casei strain. Different extraction methods were evaluated in three classes of conventional, kit-based, and combined protocols. The quantity and quality of the extracted plasmid DNA were determined by spectrophotometry, agarose gel electrophoresis, and PCR. Results revealed that the yield of the extracted plasmids differed for each protocol and conventional protocols showed higher plasmid yields. We suggested an effective, inexpensive protocol to extract plasmid DNA from the recombinant L. casei for downstream biological processes.
Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran; Department of Molecular Microbiology, Kerman Branch, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Kerman, Iran.
Mojtaba
M
Alimolaei
Department of Pathobiology, Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran. Electronic address: golchin@uk.ac.ir.
Mehdi
M
Golchin
D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification*; D007780:Lacticaseibacillus casei / Q000737:chemistry*; D010957:Plasmids / Q000737:chemistry / Q000302:isolation & purification*
D004269:DNA, Bacterial
Extraction; Lactobacillus casei; PCR; Plasmid