• Journal of Microbiology and Biotechnology
• /
• 제7권1호
• /
• pp.21-24
• /
• 1997

Chloramphenicol acetyl transferase (CAT) protein and mRNA levels in E. coli were determined following induction of a tac::cat construct by isopropyl-${\beta}$-thiogalactopyranoside (IPTG). High cat mRNA levels did not directly reflect CAT protein levels, in either shakeflask experiments or fermentations. Furthermore, concentrations of IPTG resulting in the highest levels of expression of cat mRNA, were different to those resulting in highest levels of CAT protein. The data suggest that high transcriptional activities lead to limitations at the translational level.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1995년도 제3회 추계심포지움
• /
• pp.153-155
• /
• 1995

To investigate the mechanism of the regulation of cytochrome P450IAl, the 5'-flanking region of a trout cytochrome P4501Al was cloned into the CAT basic expression vector at HindⅢ site. This trout Cytochrome P450IAl upstream DNA containing CAT construct was transfected into Hepa-1 cells .3MC treatment to hepa I cells transfected with trout P450IAl-CAT construct increased CAT protein and mRNA by 2.81 fold when it was compared with that of control. This increase CAT protein and mRNA was decreased by concomitantly treated flavonoids and aminopyrine. The level of CAT protein was 29.2-58.0% of 3MC stimulated CAT protein.

• Mycobiology
• /
• 제41권3호
• /
• pp.145-148
• /
• 2013

Vegetative growth signaling of the opportunistic human pathogenic fungus Aspergillus fumigatus is mediated by GpaA ($G{\alpha}$). FlbA is a regulator of G protein signaling, which attenuates GpaA-mediated growth signaling in this fungus. The flbA deletion (${\Delta}flbA$) and the constitutively active GpaA ($GpaA^{Q204L}$) mutants exhibit enhanced proliferation, precocious autolysis, and reduced asexual sporulation. In this study, we demonstrate that both mutants also show enhanced tolerance against $H_2O_2$ and their radial growth was approximately 1.6 fold higher than that of wild type (WT) in medium with 10 mM $H_2O_2$. We performed quantitative PCR (qRT-PCR) for examination of mRNA levels of three catalase encoding genes (catA, cat1, and cat2) in WT and the two mutants. According to the results, while levels of spore-specific catA mRNA were comparable among the three strains, cat1 and cat2 mRNA levels were significantly higher in the two mutants than in WT. In particular, the ${\Delta}flbA$ mutant showed significantly enhanced and prolonged expression of cat1 and precocious expression of cat2. In accordance with this result, activity of the Cat1 protein in the ${\Delta}flbA$ mutant was higher than that of $gpaA^{Q204L}$ and WT strains. For activity of the Cat2 protein, both mutants began to show enhanced activity at 48 and 72 hr of growth compared to WT. These results lead to the conclusion that GpaA activates expression and activity of cat1 and cat2, whereas FlbA plays an antagonistic role in control of catalases, leading to balanced responses to neutralizing the toxicity of reactive oxygen species.

• 약학회지
• /
• 제39권6호
• /
• pp.676-680
• /
• 1995

The nucleotide sequence of Xbal-Mbol fragment of pKH7, a chloramphenicol-resistant($Cm^{r}$) plasmid isolated from multidrug-resistant S. aureus SA2, has been determined. Xbal-Mbol fragment of pKH7 was found to contain two ORFs. One ORF encoded Rap and the other encoded CAT protein. The deduced amino acid sequences of Rep and CAT of pKH7 were compared to those of pUB112 and pC221. Comparisons revealed that there was one amino acid difference in CAT between pKH7 and pUB112. CAT of pKH7 exhibited 98.6% amino acid identity to that of pC221. In case of Rep proteins, a slightly lower homology of 96.4% and 86.7% in amino acid sequences was observed between pKH7 and pUB112 and between pKH7 and pC221, respectively.

• 운동과학
• /
• 제26권3호
• /
• pp.204-211
• /
• 2017

PURPOSE: This study was to investigate the effects of resistance exercise on serum inflammatory markers and CatSper 1-4 expression in testis of OLETF rats. METHODS: Male OLETF rats were divided into two groups; control group (n=12), resistance exercise group (n=12). The exercise group performed a total of 8 weeks of moderate intensity resistance exercise on a 1.35 m vertical ladder with weights secured to their tails. RESULTS: The results of this study was following; The exercise group showed a significant decreased in the inflammatory ($IL-1{\beta}$, IL-6, $TNF-{\alpha}$) levels as compared to the control group. But CRP was no significant difference between control and exercise groups. Also, CatSper 1 and 2 were significantly increased in the exercise group compared to the control group, whereas CatSper 3, 4 were no significant difference between both groups. CONCLUSIONS: This study suggests that resistance exercise training can contribute to reduce pro-inflammatory responses in whole body and it affects male reproductive function by improve sperm quality and CatSper protein expression.

• Molecules and Cells
• /
• 제26권1호
• /
• pp.87-92
• /
• 2008

We employed dual color Fluorescence Cross Correlation Spectroscopy (FCCS) to measure the interaction between PKA regulatory (RII) and catalytic subunits (CAT) in living cells. Elevation of intracellular cAMP with forskolin decreased the cross-correlation amplitude between RFP-fused RII (RII -mRFP) and GFP-fused CAT (CAT-EGFP) by 50%, indicating that cAMP elevation leads to dissociation of RII-CAT complexes. Moreover, diffusion coefficient analysis showed that the diffusion rate of CAT-EGFP was significantly increased, suggesting that the decreased RII-CAT association caused by cAMP generated free CAT subunits. Our study demonstrates that in vivo FCCS measurements and their quantitative analysis permit one not only to directly quantify protein-protein interactions but also to estimate changes in the intracellular cAMP concentration.

• Archives of Pharmacal Research
• /
• 제20권5호
• /
• pp.404-409
• /
• 1997

Trout CYP1A-CAT expression construct was generated by cloning -3.5 Kb $5^I$ flanking DNA of trout liver CYP1A gene in front of CAT gene at pCAT-basic vector. Hepa 1 cells, which are known to contain a functional arylhydrbcarbon $receptor^I$ were transfected with trout CYP1A-CAT using lipofectin. 3-Methylcholanthrene (1 nM) was added into hepa 1 cells in culture in order to examine if $5^I$ flanking DNA of trout CYP1A gene could interact with mouse transactivating factors to bring about transcription of the chloramphenicol acetyltransferase(CAT) reporter gene. The level of CAT protein was measured by CAT ELISA and the level of CAT mRNA was determined by RTPCR. The treatment of 1 nM 3-methylcholanthrene resulted in two fold increases in CAT protein as well as CAT mRNA compared to untreated control hepa 1 cells. These data indicate that arylhydrocarbon receptors of mouse hepa 1 cells are functional to activate exogenously transfected trout CYP1A-CAT construct in terms of both transcription and translation of CAT. We also examined the effect of 3-methylcholanthrene on endogenous cyplal activity in hepa 1 cell. 3-Methylcholanthrene (1 nM) treatment to hepa 1 cells trahsfected with trout CYP1A-CAT construct stimulated the level of cyp1a1 mRNA by two folds and the activity of ethoxyresorufin-O-deethylase by two fold compared to that of control cells. In this study we reported that trout CYP1A-CAT reporter gene expression construct could be expressed by 3-methylcholanthrene treatment in mouse hepa 1 cells. Thus trout CYP1A-CAT could serve as a good model to study the mechanism of regulation of CYP1A1 gene expression.

• Journal of Microbiology
• /
• 제39권3호
• /
• pp.168-176
• /
• 2001

Five different types of catalases from photosynthetic bacterium Rhodospirillum rubrum S1 grown aerobically in the dark were found in this study, and designated Catl (350 kDa), Cat2 (323 kDa), Cat3 (266 kDa), Cat4 (246 kDa), and Cat5 (238 kDa). Analysis of native PAGE revealed that Cat2, Cat3, and Cat4 were also produced in the cells anaerobically grown in the light. It is notable that only Cat2 was expressed much more strongly in response to the anaerobic condition. Enzyme activity staining demonstrated that Cat3 and Cat4 had bifunctional catalase-peroxidase activities, while Catl, Cat2, and Cat5 were typical monofunctional catalases. S1 cells grown aerobically in the presence of malate as the sole source of carbon exhibited an apparent catalase Km value of 10 mM and a Vmax of about 705 U/mg protein at late stationary growth phase. The catalase activity of Sl cells grown in the anaerobic environment exhibited a much lower Vmax of about 109 U/mg protein at late logarithmic growth phase. The catalytic activity was stable in the broad range of temperatures (30$\^{C}$-60$\^{C}$), and pH (6.0-10.0). R. rubrum S1 was much more resistant to H$_2$O$_2$in the stationary growth phase than in the exponential growth phase regardless of growth conditions. Cells of stationary growth phase treated with 15 mM H$_2$O$_2$for 1 h showed 3-fold higher catalase activities than the untreated cells. In addition, L-glutamate induced an 80-fold increase in total catalase activity of R. rubrum S1 compared with magic acid. Through fraction analyses of S1 cells, Cat2, Cat3, Cat4 and Cat5 were found in both cytoplasm and periplasm, while Catl was localized only in the cytoplasm.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 1995년도 춘계학술대회
• /
• pp.128-128
• /
• 1995

To investigate the mechanism of the regulation of cytochrome P450IA1 gene expression, ethoxyresorufin deethylase(EROD) and benzo(a)pyrene hydroxylase in B6 mouse liver, in isolated perfused rat liver system. and in B6 mouse hepatocyte Hepa-I cells were examined. In C57BL/6N mouse, 3-methylcholan- throne( 3MC ) treatment have resulted in the stimulation of EROD activity based on fluorometry by 2.79 fold comparirng with that of control. Measurement of mRNA of cytochrome P450 was carried out by either nothern blot or dot blot analysis. Findings are similar to that of studies with enzymes. Furhtermore, when RTPCR method was applied to detect mRNA in Hepa I cell and liver tissues the results were more clear. Cytochrome P450IA1 upstream DNA containing CAT construct was transfected into Hepa-1 cells. After transfection of CAT construct, 3MC and flavonoids, such as, chrysin, hesperetin, kaempferol, morin, myricetin and aminoyrine were treated. 48 Hours after treatments, cells were harvested and assayed for CAT mRNA by RTPCR. 3MC treatment to hepa I cells transfected with trout P450IA1-CAT construct increased CAT mRNA by 2.81 fold when it was compared with that of control. This increase CAT mRNA was decreased by concomitantly treated flavonoids and aminopyrine. The level of CAT protein was 29.2-58.0% of 3MC stimulated CAT protein. Results of this study suggested that RTPCR seems to be a very good method to study regulation of gene expression in liver tissue or Hepa cells.

• 한국자원식물학회지
• /
• 제9권3호
• /
• pp.203-210
• /
• 1996

더덕 뿌리 내에 함유되어 있는 항산화효소를 생육환경에 따라 조사하기 위하여 경북 서벽지역의 1 2년생 밭더덕, 야생더덕 및 논에서 재배되는 1 2년생 더덕을 채취하여 사용하였고, 부위별 비교는 각 더덕 뿌리를 upper, middle, low로 나누어서 사용하였다. 이들을 O radical caseade인 SOD, POD, CAT activity를 조사하여 다음과 같은 결과를 얻었다. 1. SOD 활성은 밭재배조건의 경우 1년생보다는 2년생이, 논재배시는 1년생이 2년생보다는 높은 활성을 나타내었으며, 야생종의 비활성이 96069u/mg protein으로 가장 높은 활성을 나타내었다. 부위별로 비교해보면 재배종은 upper>middle>lower 순으로 논재배시는 low>upper>middle 순으로 야생종은 middle>lower, upper 순으로 생육환경에 따라 차이를 나타내었다. 2. POD 비활성(u/mg protein)은 년근수가 높을수록 활성은 증가하며, 야생지역이 68 u/mg protein로 가장 높은 활성을 나타내었다. 부위별로는 middle>lower>upper 순으로 활성이 나타났으며, 서벽야생의 middle 부분이 85 u/mg protein를 나타내었다. 3. CAT 활성은 POD 활성과는 달리 년근수가 낮을수록 높게 나타났으며 밭재배지와 야생지의 활성은 비슷하며 논재배지는 상대적으로 그 활성이 낮았다. 부위별로는 upper>low>middle 순으로 upper 부위가 상당히 높은 활성을 나타내며, 서벽재배 1년생의 upper가 53.59 u/mg protein으로 가장 높게 나타났다. 4. MS1D(MS+2.4-D 1ppm)에 배양한 배양세포의 항산화효소의 활성은 CAT는 1564, POD는 30, SOD는 22200 u/mg protein로 생체에 비해 낮은 활성을 나타내었다.