• 생태와환경
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• 제38권4호통권114호
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• pp.510-517
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• 2005

선형의 남조류 아나베나는 광합성과 질소고정 능이 있으며, 이는 아마도 이 남조류가 각종 육수 환경에 잘 적응하는데 큰 역할을 했다고 볼 수 있다. 작은 patS라는 유전자는 질소 고정세포의 형성을 막으며 결과적으로 질소원이 부족한 환경에서 아나베나의 죽음을 가져온다. 본 연구는 patS 유전자 주변의 DNA 염기 서열을 분석하여 codon 활용도를 알아 본 결과 patS를 제외한 다른 유전자가 존재하지 않음을 밝혔다. patS를 포함하는 세 개의 겹치는 cosmid를 찾아서 기존의 알려진 이형세포 발달 유전자를 탐색하였으나 나타나지 않았다. 질소원의 결핍에 반응하는 patS 유전자의 발현을 Northern blot 분석과 lacZ reporter 유전자 합성 실험을 통하여 각각 전사와 번역의 단계에서 알아보았다. patS 유전자의 발현은 배지로부터 질소원이 제거된 후 12시간 내에 급격히 증가하는 것으로 나타났다.

• KSBB Journal
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• 제10권1호
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• pp.89-96
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• 1995

A rabidopsis thaliana의 trpl 변이 식물체는 uv 하에서 푸른 형광 빛을 발하며, phosphoribosyl anthranilate transferase를 encoding하는 유전자가 결여되어 있다. 이 유전자를 PATl이라고 하며, 많 은 미 생물에서 phospho ribosyl anthranilate trans ferase와 homologous하다. 트럽토판 생합성에서 이 효소가 결여되면 anthranilate가 축적되며 형광 빛 을 발하게 된다. PATl 의 유전자 조절을 알아보기 위하여, PATl 유전자의 promoter를 단계 별로 삭제하여 트립토판 변이 식물체의 형질전환과 재분화를 시도하였다. 이 러한 유전자 조작을 통하여 이 유전자의 발현 양상을 조절하는 promoter 요소와 작용을 확인할 수 있으리라고 생각된다. 또한 PAT의 항체를 사용한 Immunoassay를 통하여 형질전환체 의 단백질 양의 변화를 분석한 결과 PATl의 완전 한 promoter를 가진 pHSI07의 형질전환체는 대조구보다 2배의 단백질 양을 나타냄으로서 2쌍의 PAT 유전자가 발현되었음을 알 수 있었다. PATl 은 selection marker와 reporter 유전자로서 분자유 전학연구에 공헌할 수 였으리라고 생각된다.

• Journal of Plant Biology
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• 제39권4호
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• pp.243-247
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• 1996

Phosphoribosylanthranilate transferase (PAT) catalyzes the second step of the tryptophan biosynthetic pathway and is encoded by a single-copy gene that complements all the visible phenotypes of the tryptophan mutant (trp1-100) of Arabidopsis. The trp1-100 is blue fluorescent under UV light becuase it accumulates anthranilate. To obtain a plant with reduced PAT activity, PAT1 genes with several internal deletions in different promoter regions (pHS 101, pHS102, pHS104, pHS105, and pHS107) were induced into trp1-100 via Agrobacterium. Then, homozygous T3 plants were isolated and examined for blue fluorescence. Introduction of the PAT1 gene fusants results in the reversion of fluorescence phenotype except in the case of pHS105. These results prompted us to perform a parallel analysis of anthranilate synthase and PAT interms of the genetic complementation. A plant line carrying pHS105 gene fusant does not completely complement the blue fluorescence but it accumulates less anthranilate than trp1-100. The activity of PAT was reduced in the transgenic mutant as well. The plant carrying these constructs will add to the growing collection of molecular tools for the study of the indolic secondary metabolism.

• Fisheries and Aquatic Sciences
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• 제19권7호
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• pp.31.1-31.6
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• 2016

Background: The objective of this study was to develop an efficient selectable marker for transgenic Dunaliella salina. Results: Tests of the sensitivity of D. salina to the antibiotic chloramphenicol and the herbicide Basta$^{(R)}$ showed that cells ($1.0{\times}10^6cells/ml$) treated with 1000 or $1500{\mu}g/ml$ chloramphenicol died in 8 or 6 days, respectively, whereas D. salina cells ($1.0{\times}10^6cells/ml$) treated with 5, 10, 20, or $40{\mu}g/ml$ Basta$^{(R)}$ died in 2 days. Therefore, D. salina is more sensitive to Basta$^{(R)}$ than to chloramphenicol. To examine the possibility of using the phosphinothricin N-acetyltransferase (pat) gene as a selectable marker gene, we introduced the pat genes into D. salina with particle bombardment system under the condition of helium pressure of 900 psi from a distance of 3 cm. PCR analysis confirmed that the gene was stably inserted into the cells and that the cells survived in $5{\mu}g/ml$ Basta$^{(R)}$, the medium used to select the transformed cells. Conclusions: The findings of this study suggest that the pat gene can be used as an efficient selectable marker when producing transgenic D. salina.

• Journal of Microbiology and Biotechnology
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• 제28권12호
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• pp.2009-2018
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• 2018

Leuconostoc mesenteroides can be used to produce mannitol by fermentation, but the mannitol productivity is not high. Therefore, in this study we modified the chromosome of Leuconostoc mesenteroides by genetic methods to obtain high-yield strains for mannitol production. In this study, gene knock-out strains and gene knock-in strains were constructed by a two-step homologous recombination method. The mannitol productivity of the pat gene (which encodes phosphate acetyltransferase) deletion strain (${\Delta}pat::amy$), the fk gene (which encodes fructokinase) deletion strain (${\Delta}fk::amy$) and the stpk gene (which encodes serine-threonine protein kinase) deletion strain (${\Delta}stpk::amy$) were all increased compared to the wild type, and the productivity of mannitol for each strain was 84.8%, 83.5% and 84.1%, respectively. The mannitol productivity of the mdh gene (which encodes mannitol dehydrogenase) knock-in strains (${\Delta}pat::mdh$, ${\Delta}fk::mdh$ and ${\Delta}stpk::mdh$) was increased to a higher level than that of the single-gene deletion strains, and the productivity of mannitol for each was 96.5%, 88% and 93.2%, respectively. The multi-mutant strain ${\Delta}dts{\Delta}ldh{\Delta}pat::mdh{\Delta}stpk::mdh{\Delta}fk::mdh$ had mannitol productivity of 97.3%. This work shows that multi-gene knock-out and gene knock-in strains have the greatest impact on mannitol production, with mannitol productivity of 97.3% and an increase of 24.7% over wild type. This study used the methods of gene knock-out and gene knock-in to genetically modify the chromosome of Leuconostoc mesenteroides. It is of great significance that we increased the ability of Leuconostoc mesenteroides to produce mannitol and revealed its broad development prospects.

• 한국자원식물학회지
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• 제17권1호
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• pp.28-33
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• 2004

본 연구는 개량 포플러 현사시 3호를 재료로 제초제 저항성 유전자 PAT으로 형질전환시 킨 다음 제초제 Basta처리에 의한 효율적인 형질전환체 선발을 목적으로 시험되었다. Basta 처리에 따른 캘러스유도, 부정아유도 및 액아유도 시험을 통해 조직치사 농도는 1.0mg/L로 결정할 수 있었으며, 이 농도처리에 의해 형질전환체의 조기선발에 사용하였다. 이 농도처리로 비형질전환체는 모두 고사되었지만, 형질전환체는 정상적으로 반응을 보이며, 생육이 가능하였다 .한편, 형질전환체에 제초제를 직접 살포한 결과, 정상적 생육이 가능했고, PAT activity측정 결과에서도 양성 반응을 나타내어 현사시 3호 세포내에 PAT유전자가 정상 발현됨이 확인되었다. 이로써 현사시에 PAT유전자 도입 및 발현을 위하여 제초제 Basta 처 리와 PAT assay가 매우 효과적인 것으로 나타났다.

• 한국자원식물학회지
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• 제11권2호
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• pp.188-194
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• 1998

This experiment was conducted to introduce phosphinothricin acetyl -transferase(PAT) gene, resistant to basta and non-selective herbidide, into tobacco(Nicotiana tabacum cv.BY4). For shoot formation,tobacco leaf disks were placed on the MS medium supplemented with 2.0mg/L BA and 0.1mg/L NAA. In this medium condition, tobacco leaf disces were cocultivated with A. tumefaciens MP90 containing NPT IIand PAT resistant to kanamycin and Basta, respectively. Shoots were obtained in the medium containing antibiotics, and those were transferred to rooting medium supplemented with 0.1mg/L NAA and antibiotics. The plants obtaining roots were transplanted into soil. Phenotype of transgenic tobacco plant was mostly as normal plant. However, about 5% was abnormal plant, which did not set seeds. PCR analysis and southern blot were performed to determine transformation. As the results, it was confirmed that PAT gene was stably integrated into tobacco genome.When herbicide, basta, was sprayed to the plants confirmed by PCR, the transgenic plants showed normal growth, whereas normal plants died. Therefore, the result of this experiment show that tobacco transformation for the resistance to basta, non-selective herbicide, was successful because PAT gene was stably integrated into tobacco.

• Journal of Microbiology and Biotechnology
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• 제17권4호
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• pp.681-684
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• 2007

An immunoassay method was developed to quantitatively detect phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance (bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was overexpressed in Escherichia coli M15 (pQE3l-bar) and efficiently purified by $Ni^{2+}$ affinity chromatography. A developed sandwich enzyme-linked immunosorbent assay (S-ELISA) method (detection limit: $0.01{\mu}g/ml$) was 100-fold more sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot analysis in detecting the recombinant PAT. In real sample tests, PAT in genetically modified herbicide-tolerant (GMHT) peppers was successfully quantified [$4.9{\pm}0.4{\mu}g/g$ of sample (n=6)] by the S-ELISA method. The S-ELISA method developed here could be applied to other GMHT crops and vegetables producing PAT.

• 식물조직배양학회지
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• 제28권6호
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• pp.353-357
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• 2001

인삼의 자엽과 callus에 Biolistic system을 이용한 비선택성 제초제인 bialaphos에 대한 내성을 갖게 하는 PAT 유전자의 형질전환효율 향상 및 형질전환체의 유전분석에 관한 실험을 수행하였다. 자엽의 경우에는 형질전환율이 약했지만 callus의 경우에는 target distance 9 cm, rupture disk-macro-carrier gap distance를 1/3"로 했을 때 가장 양호한 형질전환 결과를 보였다. 형질전환된 인삼식물체에서 PAT 및 NPT 유전자의 존재 여부를 확인하기 위해서 PCR을 수행한 결과 정상 식물체서는 전혀 PCR product가 형성되지 않은 반면 형질전환체 모두에서 PAT (약 300 bp)와 NPT (약 800 bp) 유전자의 band를 확인하여 각각의 유전자가 삽입되어 PAT 및 NPT IIgene이 도입된 형질전환체임을 확인할 수 있었다. 있었다.

• 대한두경부종양학회지
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• 제22권2호
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• pp.117-122
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• 2006

Background and Objectives : Thyroid carcinoma is the sixth commonest cancer in Korea and the papillary carcinoma is the most common type(88%) of the malignant thyroid tumors. Bulky DNA adducts formed by the carcinogens are repaired by DNA repair process, but failure to repair this DNA damage can cause mutations in oncogenes and tumor suppressor genes resulting in tumor formation. The xeroderma pigmentosum group C(XPC) gene is essential for this repair procedure and the XPC-PolyAT(PAT) polymorphisms may alter DNA repair capacity(DRC) and genetic susceptibility to cancer. Subjects and Methods : In a case-control study of 113 Korean patients with pathologically diagnosed thyroid papillary carcinoma and 65 control subjects, we investigated the association between the three XPC-PAT gene polymorphisms and thyroid papillary cancer susceptibility. Results : The frequency of the variant XPC-PAT allele was lower in the cases(0.349) than in the controls (0.423), but the difference was not significant(p=0.140). Using logistic regression adjusting for age and sex, risk for thyroid papillary cancer was not increased in the XPC-PAT-/+ and XPC-PAT+/+ compared to XPCPAT-/-(adjusted overall odds ratio[95% confidence intervals;95%CI]=0.52[0.26-1.03] and 0.62 [0.22-1.75], respectively; trend test, p=0.167). Conclusion : There are no relationship between the XPC-PAT polymorphism and the risk of thyroid papillary carcinoma in Korean population. Based on our results, XPC-PAT polymorphism do not modulate genetic susceptibility to thyroid papillary cancer.