TY  - JOUR
TI  - Measurement of the Length of the Integrated Donor DNA during Bacillus subtilis  Natural Chromosomal Transformation
AU  - Serrano, Ester 
AU  - Carrasco, Begoña 
VL  - 9
IS  - 16
PY  - 2019
DA  - 2019/08/20
SP  - e3338
C1  - Bio-protocol 2019;9:e3338
DO  - 10.21769/BioProtoc.3338
UR  - https://doi.org/10.21769/BioProtoc.3338
AB  - For natural transformation to occur, bacterial cells must first develop a programmed physiological state called competence. Competence in Bacillus subtilis, which occurs only in a fraction of cells, is a transient stress response that allows cells to take up DNA from the environment. During natural chromosomal transformation, the internalized linear single-stranded (ss) DNA recombines with the identical (homologous) or partially identical (homeologous) sequence of the resident duplex. The length of the integrated DNA, which can be measured, depends on the percentage of sequence divergence between the donor (internalized) and the recipient (chromosomal) DNAs.The following protocol describes how to induce the development of competence in B. subtilis cells, how to transform them with donor DNAs representing different percentages of sequence divergence compared with the recipient chromosomal DNA, how to calculate the chromosomal transformation efficiency for each of them, and how to amplify the chromosomal DNA from the transformants in order to measure the length in base pairs (bp) of the integrated donor DNA.
KW  - Natural chromosomal transformation
KW  - Bacillus subtilis competent cells
KW  - Divergent DNA sequences
KW  - Integration length
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False