Cloning and Partial Characterization of the Bacillus thuringiensis dendrolimus T84A1 Insecticidal Crystal Protein Gene
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概要
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Pst I -fragments from three plasmid DNAs (10.0, 19.3 and 51.1 kb) of Bacillus. thuringiensis (Bt) dendrolimus T84A1 were cloned in Escherichia coli. One of transformants, which carrys a 10.22 kb Bt DNA (9.91 and 0.31 kb Pst I -segments), produced a 145 k polypeptide which has the same electrophoretic mobility as the crystal protein (CP) and reacts with anti-CP antibodies. Deletion analysis revealed that the entire CP gene resided on the 5.0 Kb Hpa I -Pst I region within the 9.91 kb Pst I -segment. Hybridization analysis showed that the cloned gene was derived from the largest plasmid and might be the only one in this Bt strain. The restriction enzyme map of the cloned segment was very similar to it of the sotto strain except the 5' -adjacent region of the CP gene.B. thuringiensis (Bt) dendrolimus T84A1のプラスミドから、殺虫性結晶蛋白質の遺伝子を大腸菌によってクローン化した。本菌は10.0, 19.3と51.1kbの3種のプラスミドを保有していた。PstI消化で生じた断片をベクターpBR322と連結して大腸菌を形質転換した。結晶蛋白質の抗体と反応し同じ分子量の蛋白質を生産する1株が得られた。これは10.22kbのBtDNAを含む組換えプラスミドをもち、欠失実験と毒素生産能分祈から5kbのHpaI-PstIに領域に遺伝子が認められた。ハイブリダイぜイション分析から、クローン化された遺伝子が最も大きいプラスミドに由来すること、本菌中唯一のものであること、その周辺に何らかの反復配列があることが明らかになった。本遺伝子部はsotto株のそれと類似していたが、5'-隣接部の変化が認められた。
- 広島大学生物生産学部,農林水産研究情報センターの論文
著者
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Nagamatsu Yasunori
Faculty Of Applied Science Hiroshima University
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永松 康徳
広島大 生物生産
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Nagamatsu Y
Department Of Applied Biochemistry Faculty Of Applied Biological Science Hiroshima University
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