Improvement of bacterial transformation efficiency using plasmid artificial modification
نویسندگان
چکیده
We have developed a method to improve the transformation efficiency in genome-sequenced bacteria, using 'Plasmid Artificial Modification' (PAM), using the host's own restriction system. In this method, a shuttle vector was pre-methylated in Escherichia coli cells, which carry all the putative genes encoding the DNA modification enzymes of the target microorganism, before electroporation was performed. In the case of Bifidobacterium adolescentis ATCC15703 and pKKT427 (3.9 kb E. coli-Bifidobacterium shuttle vector), introducing two Type II DNA methyltransferase genes lead to an enhancement in the transformation efficiency by five orders of magnitude. This concept was also applicable to a Type I restriction system. In the case of Lactococcus lactis IO-1, by using PAM with a putative Type I methyltransferase system, hsdMS1, the transformation efficiency was improved by a factor of seven over that without PAM.
منابع مشابه
Impact of heat shock step on bacterial transformation efficiency
CaCl2 treatment followed by heat shock is the most common method for artificial transformation. Here, the cells were transformed using CaCl2 treatment either with heat shock (standard protocol) or without heat shock (lab protocol) to comprehend the difference in transformation efficiency. The BL21 strain of Escherichia coli (E. coli) was being susceptible using CaCl2 treatment. Some Cells were ...
متن کاملروش سریع و مؤثر ترانسفورمسازی باکتری اشریشیاکلی
Background & Objective: Transformation of plasmid DNA into bacterial competent cells is a key technique for molecular cloning. Transformation can be achieved using either chemical or physical methods, e.g., electroporation. The rate of success in these methods depends on experience and attention to method’s details. Therefore, the higher the efficiency and quality of a transformation method, ...
متن کاملبهینهسازی بیان سم شقایق دریایی (فراگاسیاتوکسین C) در باکتری E.coli سویه BL21
Background and purpose: Sea anemones of the genus Actinia are commonly found in the intertidal zone of the northern rocky coast of Spain. Actinia fragacea is one of the most important species that secretes a toxic protein, fragaceatoxin C (FraC), for defense and hunting purposes. The aim of this study was transformation of plasmid containing the gene encoding the FraC to Escherichia coli bacter...
متن کاملGenetic Transformation of Amylase Gene to Ruminal Bacteroides Species Using Conjugation Consequence for Improvement of Rumen Enzyme
Rumen bacterial strains can potentially be manipulated to perform functions different from wild type species. The most numerous species of bacteria in the rumen and gut are species of the familyBacteroidetes, whichcan have the potential for genetic modification for enzyme production. One of the genetic manipulation of rumen bacteria can perform for production of starch digestive enzyme for the ...
متن کاملDataset of plasmid DNA extraction using different magnetic nanoparticles (MNPs)
In this dataset we integrated figures related to bacterial transformation using pBI121 plasmid and complementary analysis for magnetic nanoparticles (MNPs) characterizations. The structural map of pBI121 plasmid was drawn by Vector NTI software using the complete sequence of binary vector pBI121. Escherichia coli bacteria transformed using pBI121 plasmid and were grown on the selection media co...
متن کامل