SDS-PAGE를 이용한 Prevotella intermedia와 P. nigrescens의 감별에 관한 연구
DIFFERENTIATION OF PREVOTELLA INTERMEDIA AND P. NIGRESCENS USING SDS-PAGE
소속 상세정보
배광식/Kwang Shik Bae
KMID : 0362319970220020693
Abstract
In 1992, Prevotella intermedia was shown to be comprised of another spoecies now
known as Prevotella nigrescens. Strain ATCC 33563 is now designated the type strain
of P. nigrescens while strain ATCC 25611 is remains the type strain of P. intermedia.
The purpose of this study was to find the differences in protein profiles of P.
intermedia and P. nigrescens, using sodium dodecyl sulfate polyacrylamide gel
electrophoresis, which can be used for differentiation of those two species. A partial
amino acid sequence of the 18.6 kDa protein band, which was specific in P. nigrescens,
was also determined.
The cellular proteins were extracted from the cell pellets of pure cultures of P.
intermedia. and P. nigrescens by either sonication or being shaken contonuously for 20
min at 21℃ with 1% SDS or being boiled for 3 min with 1% SDS, SDS-Page was
performed according to the method of laemmli using either 12% (w/v) gels or 18%
(w/v) gels.
Results were as follows ; 1. The similar electrophoretic protein profiles were shown
by 3 cellular protein extraction methods for each strain.(Fig, 1 and 2).
2. The 18.6 kDa band which was specific only in P. nigrescens could be used for the
differentiation of P. intermedia. and P. nigrescens.(Fig. 1 and 2. Table 1).
3. A total of 4 different tryptic fragments from the 18.6 kDa protein were sequenced. the
resulting amino acid sequences were fragment 1.GNPVNIGGEW, 2.FNVVR, 3.
NYLTVAPY, and 4.GGDNVTTYQVLPEIGYN. By comparison to the sequences of
known proteins in the Swiss-Prot database and PIR database. 90% matching between
fragment 1 and serine hydroxymethyl transferase(P24060) in the Swiss-Prot, and 90%
matching between fragment 1 and glycine hydroxymethyl transferase(S15203) in the PIR
were shown, but the identity and function of the 18.6 kDa protein remains unknown.
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