@article{oai:niigata-u.repo.nii.ac.jp:00028474,
 author = {Haginoya, Kohsuke and Kato, Taisuke and Higuchi, Masahiro and Shitomi, Yasuyuki and Asakura, Tsuyoshi and Hayakawa, Tohru and Mitsui, Toshiaki and Hori, Hidetaka},
 issue = {2},
 journal = {新潟大学農学部研究報告, 新潟大学農学部研究報告},
 month = {Mar},
 note = {Liposomes were constructed with phosphatidylcholine (PC) and fluorescence indicator, calcein, and purified using 9-40% discontinuous sucrose density gradients centrifugation (SDGC) or gel filtration column chromatography (GFCC). Stability and sensitivity of those liposomes to insecticidal Cry1A toxins of Bacillus thuringiensis were compared each other and one purified by GFCC was shown to leak calcein by 40% of total fluorescence contents during 96 hr storage. On the other hand, liposome purified by SDGC was more stable and almost no leakage observed during the same period. Cry1Ab of B. thuringiensis weakly reacted with the liposome purified by GFCC and 2.5% of total encapsulated fluorescence was released, but Cry1Aa and Cry1Ac did not. Contrarily, liposome purified by SDGC was highly reactive with Cry1Ab and 40% of the calcein was released and 15%-release by Cry1Aa and Cry1Ac were also observed. These data clearly indicated that liposomes purified by SDGC were more stable and highly reactive to Cry toxins compared to that purified by GFCC. These liposomes are useful tool to analyze the interaction between Cry1A toxins and lipid membranes.ホスファチジルコリン(PC)を用いてリポソームを構築し、内部空間に蛍光色素、カルセインを封入した。PCリポソームをショ糖密度勾配遠心法(SDGC)あるいはゲルろ過カラムクロマトラグラフィー(GFCC)で精製し、各々の精製PCリポソームについて、溶液中での安定性とBacillus thuringiensisのCry1A殺虫毒素との反応性を比較解析した。GFCCで精製したPCリポソームは不安定で、96時間保存すると内包カルセインの40%がリポソームの外に漏出した。一方、SDGC精製PCリポソームは安定で96時間保存後もほとんどカルセインの漏出を示さなかった。殺虫毒素Cry1AbはGFCC精製PCリポソーム膜に小孔形成を示したが、カルセインの漏出は2.5%にとどまり、小孔形成率は非常に低いと思われた。またCry1AaとCry1Acは共に小孔を形成しなかった。一方、SDGC精製PCリポソームはCry1Abと反応し、40%のカルセイン漏出が検出された。Cry1AaとCry1Acとの反応に際しても15%のカルセイン漏出が検出された。これらの実験結果は、SDGC精製PCリポソームの方がGFCC精製PCリポソームよりも安定で、Cry1A毒素との反応性も高いことを示し、比重に基づきリポソームを精製することが均一な安定リポソームを得るうえで有効であることを示した。SDGC精製PCリポソームはCry1A毒素と脂質膜との相互作用を解析する上で有効である。},
 pages = {115--120},
 title = {Preparation of Stable Liposomes Using Sucrose Density Gradient Centrifugation and Their Interaction with Insecticidal Cry1A Toxins of Bacillus thuringiensis},
 volume = {57},
 year = {2005}
}