• 한국미생물·생명공학회지
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• 제16권2호
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• pp.126-130
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• 1988

토양에서 분리한 알카리 내성 Bacillus sp. YA-14 의 promoter를 Bacillus promoter probe vector인 pPL703을 이용하여 Bacillus subtilis내에 cloning 하였다. 얻어진 형질전환체 중 promoter 활성이 가장 높은 균주의 CAT 비활성은 8.07로 expression vector인 pPL708의 CAT 비활성보다 2.5배 이상 높았으며 대수 증식기가 끝난 이후에 그 활성이 급격하게 증가하였다. 재조합 plasmid내의 삽입 DNA 단편은 그 크기가 2.8kb이고 제한효소 BamHI, Sal I 인식부위가 각각 한군데 존재하였다.

• Journal of Microbiology
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• 제41권2호
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• pp.102-108
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• 2003

A 6.1 kb Sph I fragment from the genomic DNA of Pseudomonas putida SM 25 was cloned into the veetor pUC19. The open reading frame of catB was found to consist of 1,122 nucleotides. The sequence alignment of the catB gene products from different kinds of bacteria revealed an overall identity ranging from 40 to 98%. The catC gene contained an open reading frame of 96 codons, from which a protein with a molecular mass of about 10.6 kDa was predicted. The amino acids in the proposed activesite region of CatC were found to be almost conserved, including the charged residues. Since the catBC genes in P. putida SM25 were tightly linked, the could be regulated under coordinate transcription, and transcribed from a single promoter located upstream of the catB gene, as in P. putida RBI.

• Asian-Australasian Journal of Animal Sciences
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• 제22권5호
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• pp.712-720
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• 2009

The goal of this study was to elucidate the expression and segmental distribution of the glomerular cationic amino acid metabolism transporter-2 (CAT-2) and thus to improve our understanding of porcine cationic amino acid transporters and amino acid absorption. Porcine CAT-2 was cloned, sequenced and characterized. The predicted amino acid sequence of porcine CAT-2 shared 86.1% and 92.1% identity with human and mouse CAT-2A, respectively. The tissue distribution patterns and ontogenic changes of CAT-2 mRNAs were determined by real-time Q-PCR. The results showed that porcine CAT-2 was highly expressed in the heart and intestinal tract (duodenum, ileum and jejunum). In addition, the mRNA of CAT-2 was found in liver, lung, kidney, brain and muscle. Within the intestinal tract, CAT-2 mRNA was most abundant in the ileum and rarely expressed in the duodenum. In the duodenum, the levels of CAT-2 mRNA reached their peak on day 7 (p<0.05) while in the jejunum, levels were low on day 1 and 7 and increased rapidly after day 26 before peaking on days 30 and 60 (p<0.05). The levels then dramatically decreased by day 90 (p<0.05). In the ileum, levels achieved their maximum on day 30 and then decreased significantly on day 60 (p<0.05).

• International Journal of Industrial Entomology and Biomaterials
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• 제4권1호
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• pp.63-68
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• 2002

A cDNA encoding a putative member of cathepsin B of the thiol pretense superfamily was cloned from a cDNA library of the mulberry longicorn beetle, Apriona germari. Sequence analysis of the cDNA encoding the cathepsin B of A. germari (AgCatB) revealed that the 972 bp cDNA has an open reading frame of 324 amino acid residues. The deduced protein sequence of the AgCatB showed high homology with cathepsin B of the insects, Bombyx mori (47.3% amino acid identity), Helicoverpa armigera (46.6%) and Sarcophaga peregrina (45.6%), and the lowest homology with Aedes aegypti (33.2%). The AgCatB contains six disulfate bonds typical for cysteine pretenses. The three amino acid positions Cys-109, His-267, and Asn-287 which are conserved, active sites characteristic for cathepsin B, were also found. Phylogenetic analysis further confirmed that the AgCatB has a close relationship with that of B. mori, H. armigera and S. peregrina.

• Archives of Pharmacal Research
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• 제20권5호
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• pp.404-409
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• 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
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• 제34권4호
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• pp.334-340
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• 1996

The enzyme, cis,cis-muconate lactonizing enzyme has been proposed to play a key role in the $\beta$-ketoadipate pathway of benzoate degradation. A 3.2-kb EcoRI fragment termed as pRSU2, isolated from a Pseudomonas cepacia genomic library was able to complement the catB defective mutant. Several relevant restriction enzyme sites were determined within the cloned fragment. In Pseudomonas putida SUC2 carrying pRSU2, the enzyme activity was relatively higher than those of the induced or partially induced state of wild type P. putida PRS2000. It was probably due to higher expression of P. cepacia catB in P. putida PRS2000. It was probably due to higher expression of P. cepacia catB in P. putida. One possible interpretation of these results is that the catB promoter in P. cepacia is recognized within P. putida, resulting in the almost same expression level.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1996년도 한국생물과학협회 국제학술대회
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• pp.236.1-236
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• 1996

No Abstract, See Full Text

• 한국미생물·생명공학회지
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• 제16권5호
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• pp.343-347
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• 1988

토양에서 분리한 알칼리 내성 Bacillus속의 chromosomal DNA로부터 promoter를 cloning하여 선별된 재조합 plasmid p-12내 의 promoter를 subcloning을 하였다. 그 결과 cloning된 promoter 내에는 서로 다른 두 가지의 promoter가 존재하는 것을 확인할 수 있었고 이로부터 각각의 promoter를 함유한 재조합 plasmid p-l2B1, p-l2B2를 제조하였다. 또한 CAT 비활성 측정에 의해 각 promoter의 활성을 비교해 본 결과 p-l2B1의 promoter는 p-l2B2의 promoter에 비해 상대적으로 높은 활성을 가지고 있었다. CAT 비활성을 생육시기에 따라 측정해 본 결과 p-l2B1과 p-l2B2는 대수증식기 이후 활성이 급증되었으며 배지 중 첨가된 1.0%의 glucose에 의해 활성이 억제되는 효과를 받았다.

• 한국지하수토양환경학회지:지하수토양환경
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• 제19권3호
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• pp.47-55
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• 2014

Arthrobacter chlorophenolicus A6 possesses several monooxygenases (CphC-I, CphC-II, and CphB) that can catalyze the transformation of 4-chlorophenol (4-CP) to hydroxylated intermediates in the initial steps of substrate metabolism. The corresponding genes of the monooxygenases were cloned, and the competent cells were transformed with these recombinant plasmids. Although CphC-II and CphB were expressed as insoluble forms, CphC-I was successfully expressed as a soluble form and isolated by purification. The specific activity of the purified CphC-I was analyzed by using 4-CP, 4-chlorocatechol (4-CC), and catechol (CAT) as substrates. The specific activities for 4-CP, 4-CC, and CAT were determined to be 0.312 U/mg, 0.462 U/mg, 0.246 U/mg, respectively. The results of this study indicated that CphC-I is able to catalyze the degradation of 4-CC and CAT in addition to 4-CP, which is a primary substrate. This research is expected to provide the fundamental information for the development of an eco-friendly biochemical degradation of aromatic hydrocarbons.

• 한국미생물·생명공학회지
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• 제22권1호
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• pp.18-22
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• 1994

Enterococcus faecalis KBL703 has three plasmids(p703/9, p703/5 and p703/4). Within p703/5, the specific DNA region that would confer replication function(replication origin) was searched by transformation experiments. In order to use as the recipient of transformation, two plasmid-cured strainsd were made from this strain. Four recombinant DNA constructs, each containing fragment of p703/5 and CAT(chloramphenicol acetyl transferase) gene were also made. And they were used to transform the plasmid-cured strains. Only one DNA construct containing 3.6 kb SalI fragment was stably maintained as plasmid in these strains. Additional experiment using another Enterococcus faecalis strain(ATCC29212) as a recipient was successfully done and it was confirmed that this newly constructed recombinant plasmid plasimid contained the replication origin from p703/5 plamid.