• 미생물학회지
• /
• 제28권3호
• /
• pp.192-198
• /
• 1990

플라스미드 pKM101과 이의 돌연변이제 pSLA의 mutator 유전자를 subcloning하여 재조합 플라스미드 pJB200과 pJB210을 선별하였고, umuC36- uvrA6-(TK 610) 균주에 도입하여 UV와 MMS에 대해서 보호효과와 돌연변이율에 미치는 영향을 조사하였다. 재조합된 플라스마드는 UV와 MMS에 대해서 nonmutability인 umu- 균주에서 완전히 돌연변이율을 회복시켰고 보호효과를 크게 증가시켰다. 이 사실은 muc 유전자가 cloning된 재조합 플라스미드가 돌연변이원의 처리에 의해 효과적인 발현을 하며, muc 유전자 만으로도 pKM101의 기능을 나타낸다는 것을 확인하였고, pSLA의 muc 유전자가 그 효과에 었어서 높은 영향을 미쳤다. 또한 pKM101의 muc gene을 포함한 pJB210 은 recA - (JC2926) 균주에서 돌연변이 유발능에 영향을 미치지 못한 것은 muc 유전자가 recA 유전자에 의존함을 알 수 있었다.

• 환경위생공학
• /
• 제11권2호
• /
• pp.9-20
• /
• 1996

Genotoxicity/mutagenicity of organic chemicals in industrial wastewater was investigated using umu-test with a Salmonella typhimurium TA1535 strain. The tester strain was derived by introducing plasmid pSK 1002, which carried a umu C - lac Z fusion gene into S typhimurium TA1535, and tester strain in the presence microsomal activation proved to be the more sensitive maker of genotoxicity. Genotoxic responses were observed in concentrated with a blue-rayon column, from 14 plants tested. The results were as follow; 1. Genotoxic responses were observed in concentrated from nine plants(64.3%) tested. 2. The results show that genotoxic activity was particulary high in the untreated wastewaters and decreased in the treated wastewaters(35.7%) 3. No significant correlation was found between genotoxicity and water ollution indicators, such as COD and BOD.

• 미생물학회지
• /
• 제30권5호
• /
• pp.371-376
• /
• 1992

플라스미드 pKM101 과 이의 돌연변이체 pSL4 는 UV 및 MMS 에 대해 높은 치사 저항성과 돌연변이률을 나타낸다. pKM101 과 pSLA 의 mucB 유전자의 일부분과 mucA 유전자를 lacZ fusion 벡터인 pMC874 에 subcloning 시켜 플라스미드 pBH31 과 pBH30 을 선별하였고 이 플라스미드들을 $recA^{+}lexA^{-}$, $recA^{-}와lexA^{+}$, 균주에 각각 도입하였다. $recA^{+}lexA^{+}$ 균주에서 pSLA 의 muc 유전자를 포함하는 pBH30 의 $\beta$-galactosidase활성은 pKM101 의 muc 유전자가 연결된 pBH31 보다 높았고 $recA^{-}$ 와 $lexA^{-}$ 돌연변이주에서는 UV 조사의 유무에 관계없이 $\beta$-galactosidase를 유도하지 못하였지만 $recA^{-}$ 균주에서는 UV 조사를 하지 않았을 때 pBH30 의 $\beta$-galactosidase가 pBH31 보다 약간 높게 유도되었다. 이러한 결과는 pKM101 과 pSLA 의 기능적 차이가 두 플라스미드의 Muc 단백질의 구조적 차이라고 생삭할 수 있지만, muc 유전자의 조절부위가 돌연변이되너 LexA repessor 가 작용하는 부위의 변화에 의한 것이라고도 생각햐ㄹ 수 있었다. 또한, umuC-lacZ 유전자를 가지는 pBH100 를 construction 하여 umu oeron 의 발현은 UV 조사에 의해 유도되며 recA 와 lexA 유전자에 의해 조절됨을 알 수 있었다.

• 생약학회지
• /
• 제34권1호통권132호
• /
• pp.80-85
• /
• 2003

Water extract from Astragali Radix (AR) was tested for the safety using Ames, Bacillus subtilis Rec, and umu gene expression mutagenicity tests. Mutagenic activity in any assays we tested was not found. In Ames test, Salmonella typhimurium TA98 and TA 100 were used to identify mutagenic property, and the number of histidine revertants was measured. In the Recassay, Bacillus subtilis ${H-17(Rec^+)\;and\;M-45(Rec^-)}$ strains were used to test DNA damage activity. In the SOS umu test, Salmonella typhimurium TA1535 containing plasmid pSK1002 was used as a test strain, and we monitored the levels of umu operon expression by measuring the ${\beta}-galactosidase$ activity. From the results, there was no DNA damage and mutagenicity of AR. Hepatotoxicity of AR to female ICR mice was also monitored by the measurements of s-GOT, s-GPT, LDH activities after oral feeding for 15 days. AR was not shown any significant changes of s-GOT, s-GPT and LDH activities in mice sera.

• 생명과학회지
• /
• 제15권2호
• /
• pp.215-219
• /
• 2005

감궁탕의 돌연변이원성을 유무를 알아보기 위해 Salmonella typhimurium TA 98 및 TA100을 이용한 돌연변이 원성 실험에서도 감궁탕은 어느 균에서도 돌연변이원성을 나타내지 않았으며, 이는 S-9 mixture 의해 감궁탕이 대사가 된 후에도 이와 유사한 경향을 나타내었다. 또한, SOS umu test의 경우에서도 $\beta-galactosidase$활성에는 별다른 영향을 미치지 않는 것으로 보아 감궁탕은 돌연변이원성을 일으키지 않는 것으로 판정되었으며 S-9 mixture처리에 의한 대사 후에도 이와 유사한 실험결과가 나타났다. 따라서 감궁탕은 그 자체 및 대사 후에도 DNA에 별다른 영향을 미치지 못하는 비교적 안전한 생약처방으로 여겨진다.

• 한국미생물·생명공학회지
• /
• 제25권2호
• /
• pp.144-150
• /
• 1997

After DNA damage, umuDC is the only SOS operon that must be induced to promote SOS mutagenesis in Escherichia coli. The recombinant plasmid pBC401 and pBC402 were constructed to fuse the lac structural genes with promoter region of umuDC operon to induce the expression of lacZ gene by DNA damage. We transformed the plasmid pBC401 and pBC402 into E. coli MC1061, lacZ deleted strain and determined the activity of $\beta$-galactosidase for various mutagen; UV, mitomycin C (MMC), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), 4-nitroqunoline-1-oxide (NQO), ethyl methanesulfonate (EMS). The $\beta$-galactosidase activities of PBC401 and pBC402 for UV, MMC, and NQO were increased in proportion to expression time until 3 hours thereafter, the activities were constant or slightly decreased. The activities for MNNG and EMS were not so high as for UV, MMC, and NQO. When MNNG and EMS were treated, $\beta$-galactosidase activity of pBC402 was slightly lower than pBC401 but when UV, MMC, and NQO were treated in pBC402, $\beta$-galactosidase activity was slightly higher than in pBC401. Therefore, the pBC402 was better than the pBC401 in terms of sensitivity for frameshift mutagen. We suggest that the plasmid pBC401 and pBC402 are easy to detect mutagens which cause frameshift mutation rather than point mutation.

• 생약학회지
• /
• 제36권2호통권141호
• /
• pp.93-96
• /
• 2005

Mutagenicity of Gamgung-tang (GGT) was tested using in vitro S-9 mixture in vitro host-mediated assay with Salmonella typhimurium. In the previous reports, GGT was tested for the safety using Ames(-S-9), Bacillus subtilis Rec, and umu gene expression mutagenicity tests. Mutagenic activity in any assays we tested was not found. In this report, we further investigated safety of GGT after metabolic activation in vivo. Ames test with S-9 mixture and host-mediated assay with Salmonella typhimurium TA98 were used to identify metagenic property of GGT. GGT was administered 3 times with i.m. to Balb/c mice did not induced mutagenic effect in Salmonella typhimurium TA98 recovered from the liver after 3.5h with i.p. treatment. Over the entire dose range $(3{\sim}150mg/mouse)$ tested no toxicity was detected to the bacterial cells. These results suggest that there was no DNA damage and mutagenicity by GGT.