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Lee-Stadelmann, Ok Young,Lee, Seungwoo,Chung, Haejoon,Guo, Quansheng,Kim, Myungwon,Pak, Chunho,Hackett, Wesley P. 경희대학교 유전공학연구소 1991 遺傳工學論文集 Vol.3 No.-
In vitro adventitious bud formation was investigated in 3 hybrid Populus clones with varying degrees of regeneration potential. Longitudinal and transverse wounding treatment of 1.0 cm leaf midvein and petiole explants both increased the shoot regneration capacity, more effectively with transverse wounding. Use of micro-cross sections (400 um in thickness) of leaf midvein or stem internode greatly improved regeneration capacity in all 3 clones. Supplementary calcium nitrate (7.05 to 9.4 mM) with optimal BA and NAA concentrations enhanced regeneration potential of micro- cross sections of 2 clones, but not of the most recalcitrant clone. Micro-cross section are excellent explant for obtaining large numbers of uniform adventitious shoots from minimum amount of material in a short regeneration time.
TRANSFORMATION OF CITRUS BY MICROPROJECTILE BOMBARDMENT
Heo, In-Ok,Riu, Key-Zung,Han, Tae-Wan,Lee-Stadelmann, Ok Young 제주대학교 방사능이용연구소 1995 연구보고 Vol.9 No.-
The conditions for plant regeneration from tissue sections or callus cells of Citrus species and introduction of DNA by microprojectile bombardment were studied. The plantlets were sucessfully regenerated from both tissue esgments and callus cells. The direct shootings from sections of stems and cotyledons occured in two species of Citrus, especially multi-shoots were formed from stem sections of 'sambokam' on Gresshoff and Doy (GD) medium containing 1㎎/1 of BA. The plantlets could be also obtained from the embryogenic callus cells derived from unfertilized and undeveloped seeds of Citrus snshiu on Murashige and Thucker (MT) medium containing auxin. The transient expression study by using GUS gene showed that the type of vector plasmid and target distance affected the transformation efficiency. Among the tested plasmids, pBI221 gave the highest GUS score. The optimum target distance was 7 ㎝.
Optimization of Cymbidium Transformation System by the Particle Gun Techniques
Hong, Kyung-Ae,Lee-Stadelmann, Ok Young,U, Zang-Kual,So, In-Sup,Cheong, Choong-Duk 제주대학교 방사능이용연구소 1995 연구보고 Vol.9 No.-
Process of particle bombardment for efficient transformation of Cymbidium virescence rhizome microcross sections was investigated using Biolistic particle delivery system with pBI121 harboring the β-glucuronidase(GUS) and the neomycin phosphotransferaseII(NPTII), and pBI221 containing GUS. The best result was obtained from the combination of 1.11㎛ tungsten particles coated with pBI121, 77.33 ㎏/㎠ helium pressure, 6.35 ㎜ gap distance, and 3.8 ㎝ target distance. Transient expression of the reporter gene, GUS, bombarded into the rhizome microsections was observed by the histochemical assay. The selectable marker gene, NPT II, delivered by bombarding the tungsten particles coated with the plasmid DNA was identified using the polymerase chain reaction technique.
Genetic Enhancement of Cold Tolerance of Cymbidium by Introducing ω-3 Fatty Acid Desaturase Gene
So,In-Sup,Hong,Kyung-Ae,U,Zang-Kual,Lee-stadelmann,Ok-Young,Song,Sung-Jun 濟州大學校 放射能利用硏究所 1996 연구보고 Vol.10 No.-
Protocorm like bodies(PLBs) were micro-sectioned by a vibratome, and the conditions for in vitro culture were optimized. PLBs were induced from the meristem of Cymbidium side-bud and proliferated on Kyoto medium (4 ml/liter of Hyponex) supplemented with 1 mg/liter of α-naphthaleneacetic acid(NAA). The good formation of adventitious PLBs was made from 400㎛ thick micro-sectioned PLBs. The growth and proliferation of PLBs reformed was most effective in Kyoto medium containing 2 or 4 ml/liter of Hyponex and 1.0 mg/liter of NAA and the shoot formation was promoted by the addition of 1 mg/liter of benzyl adenine(BA). As transient expression of β-glucuronidase(GUS) gene could be identified in the bombarded PLB cells, the micro-sectioned PLBs were bombarded with the microprojectiles coated with the plasmid pBIVA1 containing NPT II and fad7. Now the cold tolerant transformants of reformed PLBs are being selected on the kanamycin medium.
Li, Yong Fang,Zhao, Yu Fen,So, In Sup,Lee-Stadelmann, Ok Young 한국식물학회 1999 Journal of Plant Biology Vol.42 No.3
The effects of amino acids (aa) and N-(diisopropyloxyphosphoryl)-amino acids (DIPP-aa) on cell membranes were investigated by evaluating water and methyl urea permeability. Permeability coefficients P_f and P_s were determined by standard osmotic methods for cells of Pisum sativum stem base epidermis after 20min exposure to a 5 mM solution of each aa and DIPP-aa. The P_f value of P. sativum epidermal cells (untreated controls) was 1.3±0.4×10 exp (-3)㎛ s^-1. Treat ments with the diisopropyl-oxyphosphoryl derivatives of three one charged and three polar amino acids (serine, threonine, asparagine, and aspartic acid) and unsubstituted (free) serine and threonine increased water permeability up to about two fold of the control value. Serine and threonine and their DIPP-derivatives increased methyl urea permeability (controls 1.03±0.09×10 exp (-3)㎛ s^-1) 30 to 80 percent Other amino acids and their DIPP-derivatives caused small or insignificant changes of water permeability. Only certain polar amino acids and their DIPP-derivatives increased the osmotic water and methyl urea permeation through the plasma membrane. The specificity of these molecules on plasma membranes suggests that the active amino acids (serine and threonine) and their DIPP-derivatives interact with charged membrane molecules. The relatively small changes in water and methyl urea permeability may indicate that the effective aa's and their DIPP-derivatives interact with phospholipids rather than aquaporin. A concurring alteration of water channel proteins, however, cannot excluded.