|
KMID : 1094720000050050327
|
|
Biotechnology and Bioprocess Engineering
2000 Volume.5 No. 5 p.327 ~ p.331
|
|
|
Application of LFH-PCR for the disruption of SpoIIIE and SpoIIIG of B. subtilis
|
|
Kim June-Hyung
Kim Byung-Gee
|
|
|
Abstract
|
|
|
|
The application of LFH-PCR (long flanking homology region-PCR) forBacillus subtilis gene disruption is presented. Without plasmid- or phage-vector construction, only by PCR, based on a DNA sequence retrieved fromB. subtilis genome data base, kanamycin resistance gene was inserted into two genes ofB. subtilis involved in sporulation,spoIIIE andspoIIIG. The effect of gene disruption on subtilisin expression was examined and the sporulation frequency of two mutants was compared to that of the host strain. For this purpose, only 2 or 3 rounds of PCR were required with 4 primers. We first demonstrated the possibility of LFH-PCR for rapid gene disruption to characterize an unknown functional gene ofB. subtilis or other prokaryote in the genomic era.
|
|
|
KEYWORD
|
|
|
B subtilis, LFH-PCR, functional genomics
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
 
|