• Journal of Microbiology and Biotechnology
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• 제1권1호
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• pp.22-30
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• 1991

In order to secrete the Bacillus subtilis cytidine deaminase (CDase, cytidine/2'-deoxycytidine deaminase) encoded by the B. subtilis cdd gene in E. coli by the aid of signal sequences, the cdd gene was fused in-frame to either amyE or penP signal sequences and the gene expression and CDase localization were examined. For the penP signal sequence::cdd fusion, the cdd gene with 9 amino acids truncated from the 5'-terminus was fused in-frame to the signal sequence, then the $cdd^{+}$ colonies were not occurred from the minimal plate by cdd complementation. The result suggests that 9 amino acids on the $NH_2-terminal$ of CDase have an essential function in the enzyme activity. The hybrid protein obtained by fused gene amyE signal sequence::cdd structural gene gave $cdd^{+}$ phenotype and about half of the total CDase activity was found to be secreted in the periplasm of E. coli transformant JF611/pSO202. The periplasmic CDase activity of JF611 harboring pSO52 containing the intact cdd gene was considerablely lower than that of the cells harboring pSO202 carrying the hybrid cdd gene. This suggests that the CDase was secreted to the periplasm through the cytoplasmic membrane by the aid of the amyE signal sequence in the E. coli transformant.

• 미생물학회지
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• 제29권2호
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• pp.80-84
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• 1991

We cloned and expressed hepatitis B viral core antigen (HBcAg) gene in E. coli using $P_{L}$ promoter system. For optimal expression of the gene, we undertook the studies on the effects of the distance between Shine-Dalgarno (SD) sequence and start codon, copy number of repressor gene, induction temperature, and the stability of the core antigen. The results demonstrated that the induction at 37.deg.C was more efficient than at 42.deg.C, and the 11 base pairs (bp) distance between SD sequence and start codon of HBcAg gene was more efficient than the 15 bp distance in E. coli. The copy number of cI857 repressor gene did not influence on the expression of HBcAg, and the expression level of HBcAg in mutant type (low protease activity) and wild type strains was almost the same. The produced core antigen appeared to be HBcAg not HBeAg judged by two different radioimmunoassat (RIA) kits. This result suggested that the antigen was stable in E. coli.i.

• 대한의생명과학회지
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• 제19권2호
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• pp.124-131
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• 2013

Genome-wide association studies (GWAS) have identified a number of common variants associated with serum liver enzyme homeostasis in population. In the previous study, single nucleotide polymorphisms (SNPs) in several genes have been reported to be associated with serum liver enzyme levels in European population. We aimed to confirm whether the genetic variation of SMAD2 (SMAD family member 2) gene influence the serum liver enzyme levels in Korean population. We genotyped variants in or near SMAD2 in a population-based sample including 994 unrelated Korean adult. Here, we performed association analysis to elucidate the possible relations of genetic polymorphisms in SMAD2 gene with serum liver enzyme levels. By examining genotype data of a total of 944 subjects in 5 hospital health promotion center, we discovered the SMAD2 gene polymorphisms are associated with serum liver enzyme levels. The common and highest significant polymorphism was rs17736760 (${\beta}$=3.51, P=5.31E-07) with glutamic oxaloacetic transferase (GOT), rs17736760 (${\beta}$=5.99, P=1.25E-05) with glutamic pyruvate transaminase (GPT), and rs17736760 (${\beta}$=15.68, P=9.93E-07) with gamma glutamyl transferase (GGT) in all group. Furthermore, the SNP rs17736760 was consistently associated with GOT (${\beta}$=5.25, P=1.72E-06), GPT (${\beta}$=9.97, P=1.16E-05), GGT (${\beta}$=26.13, P=3.43E-06) in men group. Consequently, we found statistically significant SNP in SMAD2 gene that are associated with serum levels of GOT, GPT, and GGT. In addition, these results suggest that the individuals with the minor alleles of the SNP in the SMAD2 gene may be more elevated serum liver enzyme levels in the Korean population.

• Asian Pacific Journal of Cancer Prevention
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• 제17권7호
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• pp.3065-3069
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• 2016

Single nucleotide polymorphism (SNP) detection has been used extensively for genetic association studies of diseases including cancer. For mass, yet accurate and more economic SNP detection we have optimized tetra primer amplification refractory mutation system polymerase chain reaction (ARMS PCR) to detect three SNPs in the cytochrome P450 2E1 (CYP2E1) gene locus; i.e. rs3813865, rs2070672 and rs3813867. The optimization system strategies used were (1) designing inner and outer primers; (2) determining of their optimum primer concentration ratios; and (3) determining of the optimum PCR annealing temperature. The tetra primer ARMS PCR result could be directly observed using agarose gel electrophoresis. The method succesfully determined three SNPs in CYP2E1 locus, the results being consistent with validation using DNA sequencing and restriction fragment length polymorphisms (RFLP).

• Parasites, Hosts and Diseases
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• 제52권4호
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• pp.429-433
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• 2014

To identify sequences of Entamoeba histolytica associated with the development of amebic liver abscess (ALA) in hamsters, subtractive hybridization of cDNA from E. histolytica HM-1:IMSS under 2 growth conditions was performed: 1) cultured in axenic medium and 2) isolated from experimental ALA in hamsters. For this procedure, 6 sequences were obtained. Of these sequences, the mak16 gene was selected for amplification in 29 cultures of E. histolytica isolated from the feces of 10 patients with intestinal symptoms and 19 asymptomatic patients. Only 5 of the 10 isolates obtained from symptomatic patients developed ALA and amplified the mak16 gene, whereas the 19 isolates from asymptomatic patients did not amplify the mak16 gene nor did they develop ALA. Based on the results of Fisher's exact test (P<0.001), an association was inferred between the presence of the mak16 gene of E. histolytica and the ability to develop ALA in hamsters and with the patient's symptoms (P=0.02). The amplification of the mak16 gene suggests that it is an important gene in E. histolytica because it was present in the isolates from hamsters that developed liver damage.

• BMB Reports
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• 제36권5호
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• pp.514-519
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• 2003

The ribonucleotide reductase (RR) 2 gene of the HSV-2 strain G was cloned, sequenced, and expressed in an E. coli cell. The RR2 gene was located on the PstI 2.4 kb fragment, which was cloned and sequenced. The ORF of the gene was 1,011 bp and its termination codon was TAG; also, the CATATAA sequence was present in the promoter of the RR2 gene. A Poly A signal sequence (AATAAA) was found in the 3'-noncoding region. The RR2 proteins that were produced in the E. coli and Vero cells were confirmed using a Western blot analysis. SDS-PAGE revealed that the molecular weights of the fusion-RR2 that was produced in the E. coli cells were approximately 24 kDa and 38 kDa in the Vero cells. The RR2 proteins were soluble. The differences in the molecular weights might be due to modifications in the Vero cells.

• 미생물학회지
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• 제28권2호
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• pp.114-119
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• 1990

효모 Saccharomyces cerevisiae의 염색체상에 존재하는 superkiller 유전자인 SKIB 유전자를 cloning 시켜 ski 변이 주내에서 발현시켰다. 이 유전자의 C-말단부위에 E. coli의 tacZ 구조 유전자를 융합시켜 효모와 E. coli의 shuttle vector인 pSR605를 제조하고 이를 효모에 형질전환 시킨 후 나타나는 $\beta$-galactosidase의 융합단백질을 확인할 수 있었다.

• 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.

• 한국가금학회지
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• 제1권1호
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• pp.5-11
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• 1973

1. 닭 우모색에 있어서 흑색 melanin이 착색을 이압하는 I gene은 8주시 체중을 억제하는 작용을 하였다. 2. Barring을 나타내는 B gene이 I gene과 E gene이 나타났을 때 8주시 체중을 더욱 억제하였다. 3. 확장 흑색인자인 E gene과 I gene이 상호작용을 할 때에 8주시 체중을 억제하나 I gene이 억제작용 하는 이상의 작용은 하지 않았다. 4. I gene의 성장률 억제작용은 8주시 체중이 900-1,100g였을 때 평균 암탉에 있어서 37g 수탉에 있어서 25g 수탉에 있어서는 40g이었다. 이 시기의 성장률에 대하여 I와 E B S.의 공동작용은 더욱 억제작용을 하였는데 890-1,070g시에 암탉에 있어서는 45g, 수탉에 있어서는 53g였다. (중략)

• KSBB Journal
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• 제19권4호
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• pp.331-334
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• 2004

재조합 E. coli를 이용한 MCL-PHA의 생산에서 fatty acid pathway로부터 PHA 생합성 전구체 물질들이 만들어진다는 사실과 함께 이에 관여하는 enzymes이 밝혀지고 있다. 본 논문에서는 protein homology search로부터 탐색된 paaG와 ydbU genes의 PHA 생합성에서의 역할을 확인하기 위하여 paaG와 ydbU gene이 각각 knock-out된 mutant E. coli strains 를 제작하였다. 제작된 mutant E. coli들은 모균주들보다 낮은 PHA 농도와 함량을 가졌으며, 이러한 결과들로부터 paaG와 ydbU는 fatty acid pathway에서 PHA synthesis의 전구체 물질들을 공급한다는 사실을 확인하였다. 또한, 새로운 FadB homologous enzyme YgfG를 탐색하였으며, ygfG gene이 overexpression된 균주와 ygfG mutant를 제작하여 PHA 합성을 실험한 결과 ygfG도 paaG와 ydbU와 유사한 역할을 한다는 사실을 밝혔다. 이러한 연구결과들은 E. coli에서의 MCL-PHA 단량체들의 합성 경로를 확인하여 효과적인 PHA 생산 균주를 제작할 수 있게 할 것이다.