Journal of the korean academy of Pediatric Dentistry (대한소아치과학회지)
- Volume 25 Issue 2
- /
- Pages.292-303
- /
- 1998
- /
- 1226-8496(pISSN)
- /
- 2288-3819(eISSN)
EVALUATING TWO METHODS FOR FINGERPRINTING GENOMES FOR STREPTOCOCCUS MUTANS IN CHILDREN : A COMPARISON WITH AP-PCR AND SOUTHERN BLOT RFLP
유전자형에 따른 Streptococcus mutans의 subtyping: Southern blot RFLP와 AP-PCR을 이용한 비교
- Jeong, Tae-Sung (Deportment of Pediatric Dentistry, College of Dentistry, Pusan National University) ;
- Kim, Shin (Deportment of Pediatric Dentistry, College of Dentistry, Pusan National University)
- Published : 1998.05.30
Abstract
The arbitrary primer polymerase chain reaction(AP-PCR) and Southern blot restriction fragment length polymorphism(RFLP) were used to genotype the cariogenic pathogen S. mutans in children. Following the morphologic chracteristics of colony on selective medium for S. mutans, total genomic DNA from 155 strains was extracted by conventional methods. Among 155 strains, 143 strains (92.3%) were confirmed S. mutans by PCR with dexA gene and 114 strains were used in this study. Three random sequence 10-base oligonucleotide primers were chosen for AP-PCR. The amplified DNA products were separated electrophoretically in a 2% agarose gel containing ethidium bromide and the banding patterns were compared among different strains. For RFLP analysis, DNA was digested with EcoRI and BamHI, separated on a 0.7 % agarose gel and transferred to a nylon membrane. The membrane was probed with a previously characterised 1.6 kilobases (kb) DNA fragment cloned from gtf B gene of S. mutans. The probe was labeled with isotope[
Keywords