• Title/Summary/Keyword: combined mutagenesis

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Isolation and Antibacterial Activity of Actinomycetes Producing Growth Inhibition Compounds Against Multi-antibiotic Resistant Pseudomonas Aeruginosa (항생제 다제내성균 Pseudomonas aeruginosa에 대한 생육저해물질 생산 방선균의 분리 및 항균활성)

  • Kang, Dong-Hee;Bae, Ho-Kyung;Kim, Hyun-Soo
    • KSBB Journal
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    • v.26 no.1
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    • pp.19-26
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    • 2011
  • Of the 500 Actinomycetes isolates obtained from soil, one isolate grown on maltose as the sole carbon source produced compound BHK-P19, which inhibited the growth of multiple drug resistant P. aeruginosa 0245. Ultraviolet radiation mutagenesis curtailed production of BHK-P19. Mutation of the BHK-P19 producer using N-methyl-N'-nitro-N-nitroso-guanidine obviated the antibacterial activity to P. aeruginosa 0245, but not towards P. aeruginosa 0225. The mixing of BHK-P19 and BHK-S5 culture extracts inhibited P. aeruginosa 0254, 0225 and 1113. The combined application of BHK-P19 culture extract and Schizandra chinensis Baillon extract inhibited P. aeruginosa 0254, 0225, 0826, 1113, 1378, 1731 and 2492. Use of various concentrations of BHK-P19 culture extract and ampicillin markedly increased antibacterial activity against multi-drug resistant P. aeruginose 1113.

A Novel Approach to Investigating Protein/Protein Interactions and Their Functions by TAP-Tagged Yeast Strains and its Application to Examine Yeast Transcription Machinery

  • Jung, Jun-Ho;Ahn, Yeh-Jin;Kang, Lin-Woo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.631-638
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    • 2008
  • Tandem affinity purification (TAP) method combined with LC-MS/MS is the most accurate and reliable way to study the interaction of proteins or proteomics in a genome-wide scale. For the first time, we used a TAP-tag as a mutagenic tool to disrupt protein interactions at the specific site. Although lots of commonly used mutational tools exist to study functions of a gene, such as deletional mutations and site-directed mutagenesis, each method has its own demerit. To test the usefulness of a TAP-tag as a mutagenic tool, we applied a TAP-tag to RNA polymerase II, which is the key enzyme of gene expression and is controlled by hundreds of transcription factors even to transcribe a gene. Our experiment is based on the hypothesis that there will be interrupted interactions between Pol II and transcription factors owing to the TAP-tag attached at the C-terminus of each subunit of Pol II, and the abnormality caused by interrupted protein interactions can be observed by measuring a cell-cycle of each yeast strain. From ten different TAP-tagged strains, Rpb7- and Rpb12-TAP-tagged strains show severe defects in growth rate and morphology. Without a heterodimer of Rpb4/Rpb7, only the ten subunits Pol II can conduct transcription normally, and there is no previously known function of Rpb7. The observed defect of the Rpb7-TAP-tagged strain shows that Rpb7 forms a complex with other proteins or compounds and the interruption of the interaction can interfere with the normal cell cycle and morphology of the cell and nucleus. This is a novel attempt to use a TAP-tag as a proteomic tool to study protein interactions.

Radioprotective Potential of Panax ginseng: Current Status and Future Prospectives (고려인삼의 방사선 방어효과에 대한 연구현황과 전망)

  • Nam, Ki-Yeul;Park, Jong-Dae;Choi, Jae-Eul
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.4
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    • pp.287-299
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    • 2011
  • Pharmacological effects of Panax ginseng have been demonstrated in cardiovascular system, endocrine secretion and immune system, together with antitumor, anti-stress and anti-oxidant activities. Modern scientific data show protective effect of ginseng against bone marrow cell death, increased survival rate of experimental animals, recovery of hematopoietic injury, immunopotentiation, reduction of damaged intestinal epithelial cells, inhibition of mutagenesis and effective protection against testicular damages, caused by radiation exposure. And also, ginseng acts in indirect fashion to protect radical processes by inhibition of initiation of free radical processes and thus reduces the radiation damages. The research has made much progress, but still insufficient to fully uncover the action mechanism of ginseng components on the molecule level. This review provides the usefulness of natural product, showing no toxic effects, as an radioprotective agent. Furthermore, the further clinical trials on radioprotection of ginseng need to be highly done to clarify its scientific application. The effective components of ginseng has been known as ginsenosides. Considering that each of these ginsenosides has pharmacological effect, it seems likely that non-saponin components might have radioprotective effects superior to those of ginsenosides, suggesting its active ingredients to be non-saponin series. These results also show that the combined effects of saponin and non-saponin components play an important role in the radioprotective effects of ginseng.

Artificial Mutation for Silkworm Molecular Breeding Using Gene Scissors (유전자 가위의 이용과 누에 분자 육종을 위한 인위적 돌연변이 유발)

  • Hong, Jeong Won;Jeong, Chan Young;Yu, Jeong Hee;Kim, Su-Bae;Kang, Sang Kuk;Kim, Seong-Wan;Kim, Nam-Suk;Kim, Kee Young;Park, Jong Woo
    • Journal of Life Science
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    • v.30 no.8
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    • pp.701-707
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    • 2020
  • Gene editing technology using the clustered regularly interspaced short palindromic repeat (CRISPR) and the CRISPR associated protein (Cas)9 has been highly anticipated in developing breeding techniques. In this study, we discuss gene scissors as a tool for silkworm molecular breeding through analysis of Bombyx mori Kynurenine 3-Monooxygenase (BmKMO) gene editing using the CRISPR/Cas9 system and analysis of generational transmission through mutagenesis and selective crossing. The nucleotide sequence of the BmKMO gene was analyzed, and three guide RNAs (gRNAs) were prepared. Each synthesized gRNA was combined with Cas9 protein and then analyzed by T7 endonuclease I after introduction into the BM-N silkworm cell line. To edit the silkworm gene, K1P gRNA and Cas9 complexes were subsequently microinjected into the silkworm embryos; the hatching rate was 18% and the incidence of mutation was 60%. The gene mutation was verified in the heterozygous G0 generation, but no phenotypic change was observed. In homozygotes generated by self-crossing, a mutant phenotype was observed. These results suggest that silkworm molecular breeding using the CRISPR/Cas9 system is possible and could be an effective way of shortening the time required.