• KSBB Journal
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• 제24권1호
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• pp.25-29
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• 2009

Saccharomyces cerevisiae의 deletion mutant를 이용하여 ydc1, ypc1, scs7, sur1, csg2, ipt1, Icb4, Icb5, dpll의 deletion이 세라마이드의 생산에 미치는 영향을 고찰하였다. 세라마이드는 ELSD가 연결된 HPLC를 통하여 분석하였으며 ${\triangle}$ydc1 mutant의 세라마이드 생산량이 6 mg ceramide/g cell로 최대량을 나타내었으며 ${\triangle}$sur1 mutant, ${\triangle}$lcb5 mutant, ${\triangle}$dpll mutant의 경우 control로 사용한 BY4742와 비슷한 세라마이드 생산량을 나타내었고, 그 외 ${\triangle}$ypc1 mutant, ${\triangle}$scs7 mutant, ${\triangle}$csg2 mutant, ${\triangle}$ipt1 mutant, ${\triangle}$lcb4 mutant는 BY4742보다 낮은 세라마이드 생산량을 나타내었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.727-730
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• 2001

Xanthomonas oryzae #5로부터 클로닝 된 CFTase 의 functional domain의 분석을 위해 CFTase의 recombinant deletion mutant를 구성하고, recombinant protein을 분리, 정제하였다. 분리, 정제한 recombinant protein의 활성을 측정한 결과 C-terminal이 deletion 된 mutant는 cyclization 반응이 소실 되었다. 이와 같은 결과로부터 CFTase의 C-terminal 은 cyclization 반응의 중요한 functional domain 이다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.528-530
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• 2002

Xanthmonas oryzae MGL21유래의 CFTase의 repeat영역을 deletion 시킨 ${\triangle}N{\triangle}C$ deletion mutant는 protein 정제결과 약 90kDa 이있으며 pH6.5, $45^{\circ}C$에서 최적 효소 반응을 하였다. 또한 Inulin과 반응시켰을 때 CFTase는 main product가 CF인데 비해 ${\triangle}N{\triangle}C$ deletion mutant는 main product가 fructooligosaccharide였다. 이러한 결과로부터 CFTase의 N말단 repeat영역과 C말단 repeat영역을 제거하였을 경우 endoinulinase와 활성이 유사하며, 유전자 크기 및 아미노산 서열도 유사함을 알 수 있었다.

• The Plant Pathology Journal
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• 제35권5호
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• pp.393-405
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• 2019

Survival factor 1 (Svf1) is a protein involved in cell survival pathways. In Saccharomyces cerevisiae, Svf1 is required for the diauxic growth shift and survival under stress conditions. In this study, we characterized the role of FgSvf1, the Svf1 homolog in the homothallic ascomycete fungus Fusarium graminearum. In the FgSvf1 deletion mutant, conidial germination was delayed, vegetative growth was reduced, and pathogenicity was completely abolished. Although the FgSvf1 deletion mutant produced perithecia, the normal maturation of ascospore was dismissed in deletion mutant. The FgSvf1 deletion mutant also showed reduced resistance to osmotic, fungicide, and cold stress and reduced sensitivity to oxidative stress when compared to the wild-type strain. In addition, we showed that FgSvf1 affects glycolysis, which results in the abnormal vegetative growth in the FgSvf1 deletion mutant. Further, intracellular reactive oxygen species (ROS) accumulated in the FgSvf1 deletion mutant, and this accumulated ROS might be related to the reduced sensitivity to oxidative stress and the reduced resistance to cold stress and fungicide stress. Overall, understanding the role of FgSvf1 in F. graminearum provides a new target to control F. graminearum infections in fields.

• Journal of Microbiology and Biotechnology
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• 제27권1호
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• pp.72-76
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• 2017

Detection of exon 19 deletion mutation in the epidermal growth factor receptor (EGFR) gene, which results in increased and sustained phosphorylation of EGFR, is important for diagnosis and treatment guidelines in non-small-cell lung cancer. Here, we have developed a simple and convenient detection system using the interaction between G-quadruplex and fluorophore thioflavin T (ThT) for discriminating EGFR exon 19 deletion mutant DNA from wild type without a label and quencher. In the presence of exon 19 deletion mutant DNA, the probe DNAs annealed to the target sequences were transformed into G-quadruplex structure. Subsequent intercalation of ThT into the G-quadruplex resulted in a light-up fluorescence signal, which reflects the amount of mutant DNA. Due to stark differences in fluorescence intensity between mutant and wild-type DNA, we suggest that the induced G-quadruplex structure in the probe DNA can report the presence of cancer-causing deletion mutant DNAs with high sensitivity.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1338-1346
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• 2006

Ansamitocin P-3 is a potent antitumor agent produced by A. pretiosum. A deletion mutant of A. pretiosum was constructed by deleting the asm2 gene, a putative transcriptional repressor. The deletion mutant showed a 9-fold enhanced ansamitocin P-3 productivity. The response surface method with central composite design was employed to further optimize the culture medium composition for ansamitocin P-3 production by the deletion mutant. The concentrations of four medium ingredients, dextrin, maltose, cotton seed flour, and yeast extract, which have been reported as major components for ansamitocin production, were optimized through a series of flask culture experiments. The optimum concentrations of the selected factors were found to be dextrin 6.0%; maltose 3.0%; cotton seed flour 0.53%; and yeast extract 0.45%. The maximum titer of ansamitocin P-3 was 78.3 mg/l with the optimized composition, about 15-folds higher than the unoptimized titer of 5.0 mg/l obtained with YMG medium.

• 한국연초학회지
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• 제23권2호
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• pp.133-140
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• 2001

PAP-I cDNA was synthesized from total RNA of Phytolacca americana leaves by RT-PCR, and then subcloned to recombinant vector pBluescript II SK-. Using PCR with primers designed in our laboratory, we could get the 9 deletion mutant PAP-I cDNA fragments. The first of the fragments was deleted by 66bp from immature N-terminal and then the rest were deleted by 90bp sequentially. Sequentially deletion mutant PAP-I cDNAs were inserted to pAc55M, on down-stream of gall promoter. Recombinant pAc55M was transformed to yeast cells, psy1 and the cells were spreaded on SC_urn-/glucose plate media. Colonies on SC_ura-/glucose plate were streaked on the same position of SC_ura-/glucose and SC_ura-/galactose plate, and we selected colonies growing on both plates, which carry non-cytotoxic deleted mutant PAP-I cDNA. We selected 4 deletion mutant PAP-I cDNAs which have not cytotoxicity.

• BMB Reports
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• 제29권5호
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• pp.411-416
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• 1996

In order to study the role of the protein shell in both iron uptake and iron core formation of ferritin, we constructed a deletion mutant of the ferritin gene and expressed the mutant gene in Escherichia coli, This mutant was obtained by introducing an amber mutation at position Pro-157 and a deletion of the 19 amino acid residues at the carboxy-terminus of the recombinant tadpole H-chain ferritin. The deleted amino acids correspond to E-helix forming the hydrophobic channel in the protein. E. coli harboring the plasmid pTHP157, which contains the deleted gene, was grown at $23^{\circ}C$ in the presence of 0.1 mM IPTG, and the induced protein appeared to be partly soluble. Nondenaturing polyacrylamide gel electrophoresis showed that the expressed mutant H-chains coassemble into holoprotein, suggesting that E-helix is not necessary for assembly of the subunits as reported for human H-chain ferritin. Its ability in iron core formation was proven in an Fe staining gel, the result disagreeing with the observation that the hydrophobic channel is necessary for iron core formation in human H-chain ferritin.

• 생명과학회지
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• 제5권4호
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• pp.162-169
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• 1995

The glpD genes encoding gly-3-p dehydrogenase is essential for the aerobic growth of E. coli on glycerol or gly-3-p. The glpE gene, the function of which is unknownm is transcribed divergently with respect to glpD gene. Expression of the adjacent but divergently transcribed glpD the glpE genes is positively regulated by the cAMP-CRP complex. In this study, for a precise investigation of the functional elements in the regulatory region for transcription activation by cAMP-CRP, deletion mutation have been introducted into the regulatory region. The effect of the deletion mutant on transcriptional regulation was tested in vivo by $\beta$-galctosidase activity. Deletion mutants in the regulatory region of glpD demonstrated that the presence of the CRP-binding site resulted in an sixfold increase in promoter activity. And also deletion mutants of glpE gene demonstrated that the presence of the CRP-binding site resulted in an eightfold increase in promoter activity. Insertion of 22 bp oligomer in the deletion mutants has shown that the CRP binding site is need for maximal expression of glpD and glpE genes. glpD and glpE gene, cAMP-CRP complex, deletion mutant, transcriptional regulation.

• 약학회지
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• 제49권3호
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• pp.198-204
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• 2005

Human foamy virus (HFV) integrase mediates integration of viral c-DNA into cellular DNA. In this process, HFV integrase recognizes its own viral DNA specifically and catalyzes insertion of viral c-DNA. In order to study catalytic domains and residues, three deletion mutants and two point mutants of HFV integrase were constructed and analyzed with respect to enzymatic activities. The C-terminal deletion mutant showed decreased enzymatic activities while the N-terminal deletion mutant lost the activities completely, indicating that the N-terminal domain is more important than the C-terminal domain in enzymatic reaction. The point mutants, in which an aspartic acid at the 164th position or a glutamic acid at the 200th position of the HFV integrase protein was changed to an alanine, lost the enzymatic activities completely. However, they were well complemented with other defective deletion mutants to recover enzymatic activities partially. Therefore, these results suggest that the aspartic acid and glutamic acid at the respective 164th and 200th positions are catalytic residues for enzymatic reaction.