• KSBB Journal
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• 제31권1호
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• pp.27-32
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• 2016

Several CoA transferases from Clostridium beijerinckii, C. perfringens and Klebsiella pneumoniae were examined for biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) in recombinant Escherichia coli XL1-Blue strain. The CB3819 gene and the CB4543 gene from C. beijerinckii, the pct gene from C. perfringens and the pct gene from K. pneumoniae, which encodes putative CoA transferase gene, respectively, was co-expressed with the Pseudomonas sp. MBEL 6-19 phaC1437 gene encoding engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 ($PhaC1_{Ps6-19}$) to examine its activity for the construction of key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli XL1-Blue expressing the phaC1437 gene and CB3819 gene synthesized poly(3-hydroxybutyrate) [P(3HB)] homopolymer to the P(3HB) content of 60.5 wt% when it was cultured in a chemically defined medium containing 20 g/L of glucose and 2 g/L of sodium 3-hydroxybutyrate. Expression of the phaC1437 gene and CB4543 gene in recombinant E. coli XL1-Blue also produced P(3HB) homopolymer to the P(3HB) content of 51.2 wt% in the same culture condition. Expression of the phaC1437 gene and the K. pneumoniae pct gene in recombinant E. coli XL1-Blue could not result in the production of PHAs in the same culture condition. However, the recombinant E. coli XL1-Blue expressing the phaC1437 gene and the C. perfringens gene could produce poly(3-hydroxybutyrate-co-lactate [P(86.4mol%3HB-co-13.7 mol%LA) up to the PHA content of 10.6 wt% in the same culture condition. Newly examined CoA transfereases in this study may be useful for the construction of engineered E. coli strains to produce PHA containing novel monomer such lactate.

• Journal of Microbiology
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• 제42권4호
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• pp.353-356
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• 2004

In a previous study, a gst gene was isolated from the fission yeast Schizosaccharomyces pombe. This gene was dubbed gstI, and was characterized using the gstI -lacZ fusion plasmid pYSH2000. In this work, four additional fusion plasmids, pYSHSDl, pYSHSD2, pYSHSD3 and pYSHSD4, were constructed, in order to carry (respectively) 770, 551, 358 and 151 bp upstream regions from the translational initiation point. The sequence responsible for induction by aluminum, mercury and hydrogen peroxide was located in the range between -1,088 and -770 bp upstream of the S. pombe gstI gene. The same region was identified to contain the nucleotide sequence responsible for regulation by Papl, and has one puta­tive Papl binding site, TTACGTAT, located in the range between $-954\~-947$ bp upstream of the gstI gene. Negatively acting sequences are located between -1,088 and -151 bp. These findings imply that the Papl protein is involved in basal and inducible transcription of the gstI gene in the fission yeast S. pombe.

• 한국미생물·생명공학회지
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• 제13권3호
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• pp.297-302
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• 1985

미생물중 urease생성능이 아주 강한 B. pasteurii의 Hind III partial digest 된 chromosomal DNA를 E. coli-B. subtilis bifunctional plasmid vector pGR 71으로 E. coli RR1 균주에 cloning 하므로써 그 urease gene을 expression시킬 수 있었다. 그러나 B. subtilis에서는 insertion DNA fragment의 deletion으로 expression되지 않았다. Cloning된 E.coli RR1 균주로부터 분리 정제한 urease gene함유 Plasmid(pGU66)의 restriction map을 작성하여 본 결과 7.1 Mdal의 insertion fragment가 삽입된 12.6Mdal의 plasmid에 Hind III, Bgl II, Xba I, Sal I등 몇 개의 cleavage site 위치를 찾을 수 있었다. Cloning된 E. coli의 urease는 periplasmic space에 많은 비율로 축적되며, 그 효소학적 성질은 donor인 B.pasteurii의 그것과 매우 유사하였다.

• 한국동물학회지
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• 제38권2호
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• pp.204-211
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• 1995

In an effort to develop a new therapeutic strategy for human gene therapy of solid ovarian tumor, we studied the expression of the p53 tumor suppressor Sene as well as regulatory subunits of cyclic AMP (cAMP)-dependent protein kinase in human ovarian carcinoma cells. Four cell lines (2774, Caov-3, SK-OV-3 and OVCAR-3) were selected for the analyses. The p53 transcript and protein were detected only in the 2774 cell line by Northern and Western Bnalysis. In the relatively fast growing cell line, SK-OV-3, the %rope 1 a regulstorv subunit (RIA of CAMP-dependent protein kinase was the highest among the four cell lines. The expression level of $RII\beta$ protein was low in the four cell lines examined. These results maw point to a direction to select the target gene(sl to be employed for gene therapy to control the ovarian cancer.

• Journal of Microbiology and Biotechnology
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• 제10권2호
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• pp.161-168
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• 2000

Plasmid pLEX3 isolated from the recombinant cosmid library of Zoogloea ramigera 115 was found to be responsible for the restoration of the rugose colony phenotype. To confirm the essential region responsible for the complementation, subclones were constructed from plasmid pLEX3 and transformed into mutant strain Z. ramigera 115SLR. The recombinant plasmids pLEX10 and pLEX11 were shown to complement the slime-forming property of Z. ramigera 115SLR. In a compositional analysis of the exopolysaccharides from Z. ramigera 115, Z. ramigera 115SLR, and Z. ramigera 115SLR harboring plasmid pLEX11, the exopolysaccharides showed a similar composition with glucose, galactose, and side chain groups. The complete nucleotide sequence of the 3.25kb genocim DNA insert in plasmid pLEX11 was determined and its analysis identified two open reading frames which could encode two proteins. The gene products derived form the two open reading frames were confirmed by and in vivo transcription using a T7-RNA polymerase. The ORF1 produced a 30 kDa protein, whereas the ORF2 was found responsible for the complementation of the morphological mutation and produced a 14 kDa protein. An in vivo gene expression of plasmid pTEX10 showed another open reading frame encoding a 50 kDa protein. The gene products form ORF1 and ORF2 are regarded as novel proteins which do not show any homology with other proteins.

• 미생물학회지
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• 제30권6호
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• pp.539-545
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• 1992

Lactobacillus casei SM-M1 의 플라스미드로부터 phospho-$\beta$-galactosidase gene 을 갖는 DNA 를 E. coli 에 클로닝한 pPLac15(13kb) 의 재조합 플라스미드를 제조하였다.(15). pPLac15 DNA 를 분리하여 제한효소로 처리하여 제한효소 지도를 작성하였다. Phospho-$\beta$-galactosidase 유전자의 발현을 높이기 위하여 lac promoter 를 가진 pUC18 의 PstI 위치에 클닝하여 pPLac18 을 제조하였으며, 이것을 다시 EcoRI 으로 절단하여 pUC 18 에 클로닝하여 얻은 pPLac23 (7.6 kb) 를 얻었다. Phospho-$\beta$-galactosidase 효소활성은 pPLac23 의 형질전환주인 E. coli SW-23 에서는 pPLac15 를 가진 형질전환주인 E. coli SW-15 보다 약 1.8 배의 효소의 활성을 나타내었으며 pPLac18 을 가진 E. coli SW-18 보다는 약간 높은 활성을 나타내었다.

• Genomics & Informatics
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• 제21권3호
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• pp.38.1-38.8
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• 2023

Non-small cell lung cancer (NSCLC) is an important cause of cancer-associated deaths worldwide. Therefore, the exact molecular mechanisms of NSCLC are unidentified. The present investigation aims to identify the miRNAs with predictive value in NSCLC. The two datasets were downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed miRNAs (DEmiRNA) and mRNAs (DEmRNA) were selected from the normalized data. Next, miRNA-mRNA interactions were determined. Then, co-expression network analysis was completed using the WGCNA package in R software. The co-expression network between DEmiRNAs and DEmRNAs was calculated to prioritize the miRNAs. Next, the enrichment analysis was performed for DEmiRNA and DEmRNA. Finally, the drug-gene interaction network was constructed by importing the gene list to dgidb database. A total of 3,033 differentially expressed genes and 58 DEmiRNA were recognized from two datasets. The co-expression network analysis was utilized to build a gene co- expression network. Next, four modules were selected based on the Z_summary score. In the next step, a bipartite miRNA-gene network was constructed and hub miRNAs (let-7a-2-3p, let-7d-5p, let-7b-5p, let-7a-5p, and let-7b-3p) were selected. Finally, a drug-gene network was constructed while SUNITINIB, MEDROXYPROGESTERONE ACETATE, DOFETILIDE, HALOPERIDOL, and CALCITRIOL drugs were recognized as a beneficial drug in NSCLC. The hub miRNAs and repurposed drugs may act a vital role in NSCLC progression and treatment, respectively; however, these results must validate in further clinical and experimental assessments.

• Asian-Australasian Journal of Animal Sciences
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• 제29권3호
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• pp.404-412
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• 2016

We hypothesized that supplementing finishing diets with palm oil would promote adipogenic gene expression and stearoyl-CoA desaturase (SCD) gene expression in subcutaneous (s.c.) and intramuscular (i.m.) adipose tissues of feedlot steers. Eighteen Angus and Angus crossbred steers were assigned to three groups of 6 steers and fed a basal diet (control), with 3% palm oil, or with 3% soybean oil, for 70 d, top-dressed daily. Tailhead s.c. adipose tissue was obtained by biopsy at 14 d before the initiation of dietary treatments and at 35 d of dietary treatments. At slaughter, after 70 d of dietary treatment, tailhead s.c. adipose tissue and i.m. adipose tissue were obtained from the longissimus thoracis muscle. Palm oil increased plasma palmitic acid and soybean oil increased plasma linoleic acid and ${\alpha}$-linolenic acid relative to the initial sampling time. Expression of AMP-activated protein kinase alpha ($AMPK{\alpha}$) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) increased between the initial and intermediate biopsies and declined thereafter (p<0.03). SCD gene expression did not change between the initial and intermediate biopsies but declined by over 75% by the final period (p = 0.04), and G-coupled protein receptor 43 (GPR43) gene expression was unaffected by diet or time on trial. Soybean oil decreased (p = 0.01) $PPAR{\gamma}$ gene expression at the intermediate sample time. At the terminal sample time, $PPAR{\gamma}$ and SCD gene expression was less in i.m. adipose tissue than in s.c. adipose tissue (p<0.05). $AMPK{\alpha}$ gene expression was less in s.c. adipose tissue of palm oil-fed steers than in control steers (p = 0.04) and CCAAT enhancer binding protein-beta ($CEBP{\beta}$) gene expression was less in s.c. and i.m. adipose tissues of palm oil-fed steers than in soybean oil-fed steers (p<0.03). Soybean oil decreased SCD gene expression in s.c. adipose tissue (p = 0.05); SCD gene expression in palm oil-fed steers was intermediate between control and soybean oil-fed steers. Contrary to our original hypothesis, palm oil did not promote adipogenic gene expression in s.c. and i.m. adipose tissue.

• 대한바이러스학회지
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• 제26권1호
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• pp.131-137
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• 1996

Hyphantria cunea nuclear polyhedrosis virus p10유전자와 프로모터의 염기서열을 결정하였고, p10단백질의 아미노산 서열을 유도했다. pBP10재조합클론 (Cha et. al., 1991)에 삽입이 되어있는 p10유전자의 염기서열을 결정한 결과 p10유전자의 ORF는 285 bp였고, p10단백질은 95개의 아미노산으로 구성 되었으며, 분자량은 10.26 kDa이었다. 프로모터내에는 TATA box와 전사개시부위인 TAAG 염기가 발견되었다. poly (A) signal부위인 AATAAA염기서열은 3'-말단상류의 65염기부위에 위치했다. p10단백질의 N-말단은 소수성이었으며, C-말단은 고도로 친수성이었다. p10단백질에는 cysteine, histidine, tryptophane, tyrosine, glutamine, asparagine잔기가 없었다.

• BMB Reports
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• 제33권2호
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• pp.166-171
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• 2000

Thioredoxin is a multifunctional protein that is ubiquitous in microorganisms, animals and plants. Previously, the expression of the Escherichia coli thioredoxin gene (trxA) was found to be negatively regulated by cAMP. In the present study, the effect of temperature on the expression of the E. coli trxA gene was investigated. In order to examine the temperature effect, the fusion plasmid pCL70 that harbors the E. coli trxA P1P2 promoter was used. The other two fusion plasmids, pJH3 and pMH521 that were constructed in different vectors which harbor the E. coli trxA P2 promoter, were also used. When the E. coli strain MC1061/pCL70 was grown in a rich medium at $25^{\circ}C$, $34^{\circ}C$ and $42^{\circ}C$, the cells grown at $42^{\circ}C$ gave the highest $\beta$-galactosidase activity. The E. coli MC1061/pJH3 and MC1061/pMG521 cells showed increased $\beta$-galactosidase activity after the shift of the culture temperature to $42^{\circ}C$. The wild-type trxA gene of the E. coli MC1061 cells produced much higher thioredoxin activity at the higher temperature. These results support the conclusion that the E. coli trxA gene is regulated in a temperature-dependent manner. Especially the expression from its P2 promoter appeared to be sensitive to temperature.