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- 저자 : ( Shamim Amed ) (검색결과 1 건)
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Confirmation of Green Fluorescent Protein Transformants using PCR
전선영 ( Sunyoung Jeon ),( Denver Walitang ),( Shamim Amed ),( Aritra Roy Choudhury ),사동민 ( Tongmin Sa ) 한국환경농학회 2017 한국환경농학회 학술대회집 Vol.2017 No.-
Plant growth promoting bacteria (PGPB) have beneficial effects on plant growth via an enhancement of the nutrient status of their host plant. The means by PGPB enhance the nutrient status of host plants can be categorized into five areas: biological N2 fixation, increasing the availability of nutrients in the rhizosphere, inducing increases in root surface area, enhancing other beneficial symbioses of the host, and combination of modes of action. These bacteria have plant growth promoting traits such as 1-aminocyclopropane-1-carboxylate (ACC) deaminase, indole acetic acid (IAA) production, salicyclic acid, phosphate solublization, siderophore production. In endophytic relationships, PGPB actually reside inside the host plant. PGPB maybe found in the surface of the root, stems or leaves. To find the location of PGPB in plant, Green Fluorescent Protein from jellyfish Aequoria victoria used as a valuable molecular marker which can be expressed in many organisms. GFP transformants emitting fluorescence were suitable for studying plant colonization under environmental condition. This study aims to assess the process of transformation of GFP insert into bacteria by PCR amplification. For the development of gfp mutant, triparental mating was carried out in Pseudomonas frederiksbergensis OB145, Pseudomonas umsongensis OB243, Pseudomonas umsongensis OB133, Pseudomonas moorei OR108, Pantoea sp CBMB55, Brevibacterium iodium RS16 and Burkholderia pyrrocinia CBPB-HOD using donor E.coli strain pFAJ1820 and helper E.coli strain pRK2013. Antibiotic resistance is used as a selection marker to ensure that bacteria in a culture contain a particular plasmid. Transfer of plasmid pFAJ1820 into these strains was confirmed through PCR amplification with specific primer. Band had been confirmed by using Gel-doc. In this study, plant growth promoting bacteria were successfully checked for GFP transformation. Confocal microscopy will be used to confirm effective colonization and intercellular localization of bacteria strains in plants.
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