http://chineseinput.net/에서 pinyin(병음)방식으로 중국어를 변환할 수 있습니다.
변환된 중국어를 복사하여 사용하시면 됩니다.
Vibrio vulnificus ATCC 27562의 16S rRNA 유전자의 PCR과 제한효소절단 방식
허문수,정초록 한국생명과학회 1998 생명과학회지 Vol.8 No.2
A pair of designed primers (sequences from Gene Bank) amplified 16S fRNA gene of V. vulnificus within polymerase chain reaction (PCR) machine. This PCR product is about 1.3kb DNA fragment. Six enzymes (BamH I, Alu I, Sau3A I, Hind III, Sal I, Sma I) were used for restriction pattern analysis of amplified 16S rRNA gene of V. vulnificus ATCC 27562. Digested fragments are resolved by 3% agarose gel. BamH I did not show digested fragment so, there was no cutting site of BamH I in PCR product. Alu I produced three small fragments from 400 bp to 200 bp. Sau3A I produced three fragments larger than Alu I from 70 bp and 500 bp. One of fragments of Sal I was same with 500 bp of Hind III fragment and the other was 750 bp. Sma I showed two fragments of 800 bp and 470 bp. The profile of digested fragments of 16S rRNA of V.vulnificus ATCC 27562 will may be able to use standard profile for identification of V. vulnificus. Vibrio unlnificus ATCC 27562의 16S rRNA 유전자를 PCR법과 제한효소절단법으로 분석 하여 얻은 결과는 다음과 같다. 1. 고안된 한쌍의 primer로 PCR을 시행하여 얻은 산물은 약 1.3Kb 였다. 2. PCR산물을 여섯가지의 제한 효소로 절단하여 얻은 단편들은 아래와 같다. BamH I: 어떤 restriction fragment도 만들지 않았다. Alu I : 약 400bp와 200bp의 두가지 fragment를 생산하였다. Sau3A I : 약 70bp에서 450bp 사이에 세가지 fragment를 생산하였다. Hind III : 약 800bp와 500bp의 약간 큰 두가지 fragment를 생산하였다. Sal I : 약 500bp와 750bp의 두가지 fragment를 생산하였다. Sma I : 약 800bp와 470bp의 두가지 fragment를 생산하였다.
열 및 에탄을 shock 이 Virio vulnificus 의 막단백질에 미치는 영향
허문수,정초록 한국어병학회 1999 한국어병학회지 Vol.12 No.2
V. vulnificus ATCC 27562균주의 배양 온도를 47℃, 20분간 및 6% 에탄올. 10분간으로 반응시켰을 때 SDS-PAGE상에서 새로운 16가지의 heat shock protein(hsps)과 10가지의 ethanol shock protein이 나타났다. Lethal temperature에 노출하기전에 미리 열 충격을 가한 경우 thermo tolerance가 유도되었다. 균체면역에 의해 생성된 항혈청과 열 충격 세포에서 분리된 막단백질과의 ELISA에서는 Outer Membrane Protein(OMP)에서 높은 면역반응을 나타냈으며 western blotting으로는 Inner Membrane Protein(IMP)에서는 62kDa, OMP에서는 69 kDa단백이 높은 면역원성을 나타냈다. ethanol 충격 반응에서는 IMP에서는 48 kDa, OMP에서는 오직 major밴드에서만 면역반응성이 확인되었다. anti-V. vulnificus혈청에 대한 균체 응집시험에서는 열 충격 반응 후의 균체가 정상 균체에 비해 응집반응성이 높았다. New sixteen heat shock proteins (Hsps) and ten ethanol shock proteins were appeared on the analysis with SDS-PAGE when cultivation temperature for the Vibrio vulnifrcus ATCC 27562 strain was shifted-up to 42℃ from 30℃ for 20 mins and treated with of 6% ethanol for 10 mins, respectively. Even the induction of thermotolerance in V. vulnificus was coincided with the induction of Hsps if the pre-shock was adjusted to thermal temperature. Outer membrane proteins (OMPs) that were purified from the membrane of cells after heat shock showed more immunodominant pattern to the immunized rabbit anti-V. vulnificus O serum in enzyme-linked immunosorbent assay (ELISA). On the western immunoblot analysis it was confirmed that both 62 kDa IMP and 69 kDa OMP in the Hsps and 48 kDa IMP a major OMP in the ethanol shock proteins were reacted with rabbit anti-V. vulnificus O sera. Agglutination titer of the heat shocked V. vulnificus with rabbit anti-V. vulnificus O serum was higher than that of the untreated bacteria.
허문수,정초록,장경립 한국생명과학회 1999 생명과학회지 Vol.9 No.4
Baculovirus(WSBV) was isolated from infected Penaeide was collected from shrimp farm at southern sea of Korea from 1993 to 1995. The Infectious virus was purified and used for diagnosis of infected shrimp. Anti-viral serum were used for immunological detection as enzyme linked immunoabsorbent assay(ELISA) and indirect fluorescent antibody technique(IFAT). In IFAT, stomach, lymphoid organ and antenae gland of infected shrimp showed fluorescent reaction. In ELISA, tissues of spontaneously infected shrimp appeared higher O.D. values than in artificial infected shrimp. Primer set was constructed from sequence of 420bp of cloned Baculovirus(WSBV) genome. Specific band for infected shrimp was detected in Polymerase chain reaction(PCR)
허문수,Heo Moon-Soo 한국생명과학회 2005 생명과학회지 Vol.15 No.2
대하 새우양식장의 대하에 흰반점 증상을 나타내는 원인바이러스는 막대형의 이중막을 가지고 있었으며, 전자 현미경 관찰 결과 평균 크기가 $250\~300\times70\;nm$였고, 조직학적 병변은 위상피 등에서 핵이 비대해지는 것이 관찰되었다. 공격실험에서는 건강체의 새우에 많은 누적 폐사율을 보였다. 원인 바이러스 단백질은 21개의 밴드를 보였으며, 핵산 분석 결과 total 분자량은 114 kb로 나타났다. The virions of causative virus for white spot syndrome in cultured Fresh shrimp, Penaeus chinensis were rod-shaped, double envelope. An average size of the virion was 70 nm in diameter and $250\~300$ nm in length. Histopathological test of affected stomach, heart, and lymphoid organ revealed nuclear hypertrophy. Infectivity trials carried out by injection and feeding with purified virus revealed high cumulative mortality to healthy shrimp. The twenty one different protein species were detected in the analysis of virion. The length of total DNA from the purified virus particles were detected as a single band, double-stranded DNA molecule of approximately 114 kb.