• 한국환경성돌연변이발암원학회지
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• 제16권2호
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• pp.97-102
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• 1996

In this study, we have quantified genotype frequency of the cytochrome P450 (P450) 2E1 which codes for the P450 enzyme primarily responsible for the metabolic activation of carcinogenic nitrosamines and low molecular organic solvents, in Korean population by using PCR and RFLP at two sites previously associated with some cancers; a PstI and RsaI RFLP in the transcriptional regulatory region of the human P450 2E1 gene. The genotype frequencies of homozygous wild type (PstI site-absent) and heterozygous mutant type (PstI site-present) in PstI RFLP were 0.70 and 030, respectively. The homozygous mutant type in Pstl RFLP was not observed in Korean population. The genotype frequencies of homozygous wild type (RsaI site-present), heterozygous mutant type (RsaI site-absent), and homozygous mutant type in RsaI RFLP were 0.71, 0.26, and 0.03, respectively.

• 한국환경과학회지
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• 제1권2호
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• pp.99-103
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• 1992

The bacteriophages lytic for Vibrio furnissi, Vibrio furniulis and Vibrio parahemolyticus were isolated from fish gills and shellfish. Nucleic acid of bacteriophage was prepared and restriction endonuclease profile was compared. All isolates contained deoxyribonucleic acid. V. fumissi bacteriophage from fish gills showed 2 bands with Bgl II, 1 with Pst, 3 with Hind III, 1 with Bm HI and 2 with EcoR I. V Puuialis phage represented 7 fragments with Bgl II, 1 with Pst, 4 with Hind III, and 2 with EcoR I. V parhemolyticn produced 13 sites with Hind III and 4 sites with EcoR I. The fragment types were varied depending on the phage isolation. All three phages were digested with Hind III and EcoR I with different sizes. V furnissi phage were digested with 5 different restriction enzymes. Key words: Bacteriophage, Vibrio furnissi, Vibrio fluvialis, Vibrio pnrahemolyticus, Deoxyribonucleic acid, Pst, Bam HI, Hind III, EcoR I, Bgl II.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7377-7381
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• 2014

CYP2E1 PstI polymorphism G-1259C (rs3813867) genotype distributions vary significantly among different populations and are associated with both diseases, like cancer, and adverse drug effects. To date, there have been limited genotype distributions and allele frequencies of this polymorphism reported in the three major indigenous ethnic groups (KadazanDusun, Bajau, and Rungus) in Sabah, also known as North Borneo. The aim of this study was to investigate the genotype distributions and allele frequencies of the CYP2E1 PstI polymorphism G-1259C in these three major indigenous peoples in Sabah. A total of 640 healthy individuals from the three dominant indigenous groups were recruited for this study. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) at G-1259C polymorphic site of CYP2E1 gene was performed using the Pst I restriction enzyme. Fragments were analyzed using agarose gel electrophoresis and confirmed by direct sequencing. Overall, the allele frequencies were 90.3% for c1 allele and 9.7% for c2 allele. The genotype frequencies for c1/c1, c1/c2 and c2/c2 were observed as 80.9%, 18.8%, and 0.3%, respectively. A highly statistical significant difference (p<0.001) was observed in the genotype distributions between indigenous groups in Sabah with all Asian and non-Asian populations. However, among these three indigenous groups, there was no statistical significant difference (p>0.001) in their genotype distributions. The three major indigenous ethnic groups in Sabah show unique genotype distributions when compared with other populations. This finding indicates the importance of establishing the genotype distributions of CYP2E1 PstI polymorphism in the indigenous populations.

• Asian Pacific Journal of Cancer Prevention
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• 제13권10호
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• pp.4915-4921
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• 2012

Published data on any association between the CYP2E1 RsaI/PstI (c1/c2) polymorphism and liver cancer risk among east Asians are inconclusive. The aim of this Human Genome Epidemiology (HuGE) review and meta-analysis was to derive a more precise estimation of the relationship. A literature search of Pubmed, Embase, Web of science and CBM databases from inception through July 2012 was conducted. Twelve case-control studies were included with a total of 1,552 liver cancer cases and 1,763 healthy controls. Crude odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess the strength of association under five genetic models. When all the eligible studies were pooled into the meta-analysis, the results showed that the c2 allele and the c2 carrier (c2/c2 + c2/c1) of RsaI/PstI polymorphism were associated with decreased risk of liver cancer among east Asians (c2 vs. c1: OR = 0.75, 95%CI: 0.59-0.95, P = 0.016; c2/c2 + c2/c1 vs. c1/c1: OR = 0.76, 95%CI: 0.58-1.00, P = 0.050). In the stratified analysis by country, significant associations were observed between RsaI/PstI polymorphism and decreased risk of liver cancer among the Chinese population (c2 vs. c1: OR = 0.70, 95%CI: 0.54-0.91, P = 0.007; c2/c2 + c2/c1 vs. c1/c1: OR = 0.72, 95%CI: 0.54-0.95, P = 0.020), but not among Japanese and Korean populations. Results from the current meta-analysis indicates that the c2 allele of CYP2E1 RsaI/PstI (c1/c2) polymorphism may be a protective factor for HCC among east Asians, especially among China populations.

• Archives of Pharmacal Research
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• 제19권6호
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• pp.450-455
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• 1996

The genomic DNA was prepared from trout liver which was treated with 3-methycholanthrene, and cloned into lambda EMBL3 at BamHl site. The genomic library was constructed via infections of these recombinant phages into E. coli K802, and screened by the most $5^I$-portion of trout CYP1A1 cDNA. After the screening of $10^9$ clones of the amplified library, 12 positive clones were isolated, and subjected to further screenings. The results of southern blot hybridization of genomic DNA prepared from the positive clone showed the presence of a single gene of CYP1A1, and 3.5 Kb PstI fragment that hybridizes with the most $5^I$-region DNA of CYP1A1 cDNA. The restriction map of PstI fragment was determined by the restriction digestion with various enzymes. The nucleotide sequence of the upstream genomic DNA of CYPIAI was determined by DNA sequencing of exonuclease III unidirectionally deleted PstI fragment DNA using $[^{35}/S]$dATP. This paper presented the upstream genomic DNA of CYP1A1 contained a part of coding region which was about 351 base pairs (from ATG to PstI site at 3563).

• 한국미생물·생명공학회지
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• 제18권6호
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• pp.647-652
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• 1990

Bacillus thuringiensis subsp. kurstaki HD1의 내 독소생산과 내독소단백질 유전자의 클로닝에 관한 연구를 하였다. 상기 균주는 아포생성기간 중에 이중피라미드형의 내독소를 생산하였고, 크리는 약 $2.9\times 1.0 \mu m$이었다. 상기 균주는 약 10개의 플라수미드 DNA를 가지고 있었으며, 플라스미드의 분자량의 범위는 2.1에서 80kilobases였다. 플라스미드 73kb, BamHI 절단 29Kb DNA 단편과 PstI 절단 7.9Kb DNA는 Probe DNA와 혼성화되었다. PstI 7.9Kb DNA를 추출하여 운반체인 pBR322 운반체의 PstI 절단부위에 삽입하여 클로닝한 후에 E.coli HB101 균주에 형질전환하였으며, 이 클로운을 pKL-20-1로 명명했고 이 형질전환체는 Bombyxmori 유충을 치사시키는 독소물질을 생산하였다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권12호
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• pp.4977-4982
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• 2014

Background: Cytochrome P450 2E1 (CYP2E1) might be involved in the development of bladder cancer. However, previous studies of any association between CYP2E1 RsaI/PstI polymorphism and bladder cancer risk have yielded conflicting results. In this study, we performed a more precise estimation of the relationship by a meta-analysis based on the currently available evidence from the literature. Method: To assess the effect of CYP2E1 RsaI/PstI polymorphism on bladder cancer susceptibility, a meta-analysis of 6 available studies with 1,510 cases and 1,560 controls were performed through Feb 2014. Summary odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were used to estimate the strength of association for CYP2E1 RsaI/PstI polymorphism under different genetic models. Results: When available studies were pooled into the meta-analysis, we found that the C1C2 and C2C2 genotypes of CYP2E1 RsaI/PstI polymorphism significantly decreased bladder cancer risk under different genetic models (heterozygote: OR=0.766, 95%CI=0.613-0.957, $P_{OR}$=0.019; homozygote: OR=0.51, 95%CI=0.303-0.858, $P_{OR}$=0.011; dominant: OR=0.733, 95%CI=0.593-0.905, $P_{OR}$=0.004; recessive: OR=0.565, 95%CI=0.337-0.947, $P_{OR}$=0.030). Subgroup analysis indicated that C2C2 genotype was significantly associated with decreased bladder cancer risk under the homozygote genetic model in Caucasians. There was no evidence of heterogeneity or publication bias. Conclusions: The current meta-analysis suggested that the CYP2E1 RsaI/PstI polymorphism might be associated with bladder cancer susceptibility, especially in Caucasians. Further studies are needed to validate the above conclusion.

• Asian Pacific Journal of Cancer Prevention
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• 제15권13호
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• pp.5411-5416
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• 2014

Background: A number of studies have reported relationships of CYP2E1 RsaI/PstI polymorphisms with susceptibility to lung cancer in Chinese population. However, the epidemiologic results have been conflictive rather than conclusive. The purpose of this study was to address the associations of CYP2E1 RsaI/PstI polymorphisms with lung cancer risk in Chinese population comprehensively. Materials and Methods: Systematic searches were conducted in the PubMed, Science Direct, Elsevier, CNKI and Chinese Biomedical Literature Databases. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to estimate the strength of association. Results: Overall, we observed a decreased lung cancer risk among subjects carrying CYP2E1 RsaI/PstI c1/c2 and c1/c2+c2/c2 genotypes (OR=0.76, 95%CI: 0.64-0.90 and OR=0.78, 95%CI: 0.66-0.93, respectively), as compared with subjects carrying the c1/c1 genotype. In subgroup analysis, we observed a decreased lung cancer risk among c1/c2 carriers in hospital-based studies (OR=0.81, 95%CI: 0.68-0.98) and among carriers with c1/c2 and c1/c2+c2/c2 genotypes in population-based studies(OR=0.57, 95%CI: 0.42-0.79 and OR=0.58, 95%CI: 0.43-0.79, respectively), as compared with subjects carrying the c1/c1 genotype. Limiting the analysis to studies with controls in Hardy-Weinberg equilibrium (HWE), we similarly observed a decreased lung cancer risk among c1/c2 and c1/c2+c2/c2 carriers (OR=0.73, 95%CI: 0.60-0.88 and OR=0.73, 95%CI: 0.60-0.88, respectively), as compared with c1/c1. Conclusions: Our results suggested that CYP2E1 RsaI/PstI c1/c2 and c1/c2+c2/c2 variants might be a protective factor for developing lung cancer in Chinese population. Further well-designed studies with larger sample size are required to verify our findings.

• Journal of Ginseng Research
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• 제19권1호
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• pp.56-61
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• 1995

Molecular cloning and restriction mapping on ATPase $\alpha$-subunit gene (atpA) were carried out to obtain genomic information concerned with the gene structure and organization in Korean ginseng mitochondria. Two different clones containing the homologous sequence of atpA gene were selected from SalI and PstI libraries of mitochondrial DNA (mtDNA) of Korean ginseng. The sizes of mtDNA fragments inserted in SalI and PstI clones were 3.4 kb and 13 kb, respectively. Southern blot analysis with [$^{32}P$] labelled Oenothera atPA gene probe showed that atpA gene sequence was located in 2.0 kb XkaI fragment in PstI clone and in 1.7 kb XbaI fragment in SalI clone. A partial sequening ascertained that the SalI clone included about 1.2 kb fragment from SalI restriction site to C-terminal sequence of this gene but about 0.3 kb N-terminal sequence of open reading frame was abscent. The PstI fragment was enough large to cover the full sequence of atpA gene. The same restriction pattern of the overlapped region suggests that both clones include the same fragment of atiA locus. Data of Southern blot analysis and partial nucleotide sequencing suggested that mtDNA of Korean ginseng has a single copy of atpA gene. Key words ATPase a-subunit, mitochondrial DNA, Panax ginseng.

• 한국미생물·생명공학회지
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• 제19권6호
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• pp.575-582
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• 1991

글루타치온 생산증대를 도모하기 위하여 글루타치온 합성에 관여하는 효소들인 GSH-I 및 GSH-II 유전자들 pBR322 벡터에 클로닝하였다. gshI 유전자를 클로닝하기 위하여 GSH-I 효소활성이 결여된 GS903 균주를 분리하였다. E. coli K-12 염색체 DNA로부터 분리된 3.6Kb PstI DNA 절편을 pBR322 벡터에 클로닝하였다. gshII 유전자는 2.2Kb PstI-BamHI DNA 절편내에 존재하며 이 절편을 pUC13 벡터에 클로닝하였다. 플라스미드의 copy number와 gsh 유전자들을 포함하고 있는 삽입 DNA의 크기의 차이에 의한 gsh 유전자들의 발현 정도를 조사하기 위하여 pGH1090, pGH101, pGH200, pGH201, pLF4, pLF6 그리고 pGH300같은 여러 플라스미드를 사용함으로써 gshI 유전자의 발현이 의해 2배이상 증가하였다. 그러나 벡터 플라스미드내에 존재하는 gsh 유전자들을 포함하는 삽입 DNA의 크기의 차이에 의해서는 gsh 유전자들의 발현이 영향을 받지 않았다.