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Ferritin acts as a target site for the snowdrop lectin （GNA） in the midgut of the cotton leafworm Spodoptera littoralis

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Insect Science 2008年第6期15卷 513-519页

作者： Amin Sadeghi Guy Smagghe Paul Proost Els J.M. Van Damme Laboratory of Agrozoology Department of Crop Protection Laboratory of Biochemistry and Glycobiology Department of Molecular Biotechnology Faculty of Bioscience Engineering Ghent University Ghent Rega Institute Laboratory of Molecular Immunology Kathofieke Universiteit Leuven Minderbroedersstraat 10 B-3000 Leuven Belgium

The snowdrop lectin GNA （galanthus nivalis agglutinin） has been shown to possess insecticidal activity to a range of economically important insect pests. However, the precise mechanism of insecticidal action of GNA against insects remains unknown. In this investigation, we attempted to purify and identify receptor（s） responsible for binding of GNA in the larval midgut of a major lepidopteran pest （the cotton leafworm, Spodoptera littoralis） to better understand its mode of action. Therefore, cytoplasmic as well as membrane proteins from 800 larval midguts were chromatographed on a column with immobilized GNA. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the proteins eluted from the GNA column followed by sequencing of the GNA-binding proteins and BLAST analyses revealed that the N-terminal sequences of a 24 kDa polypeptide purified from the cytoplasmic and membrane protein fraction revealed sequence similarity to sequences encoding heavy chain homologs of ferritin from Manduca sexta （76% sequence identity）, Calpodes ethlius （80% sequence identity） and Bombyx mori （61% sequence identity）. Furthermore, the N-terminal sequence of a 31 kDa polypeptide from the membrane protein fraction showed sequence similarity to a light chain homolog of ferritin from Manduca sexta （88% sequence identity）.

关键词： ferritin galanthus nivalis agglutinin GNA Lepidoptera midgut receptors Spodoptera littoralis

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Study on the Obtaining of Transgenic Wheat with GNA Alien Gene by Biolistic Particle

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Agricultural Sciences in China 2002年第1期1卷 25-29页

作者： XU Qiong-fang, LI Lian-cheng, CHEN Xiao, TIAN Fang, MA You-zhi, YE Xing-guo, ZHANG Zeng-yan, XU Hui-jun and XIN Zhi-yong( Key Laboratory of Agricultural Ministry for Crop Genetics and Breeding, Institute of Crop Breeding and Cultivation ,Chinese Academy of Agricultural Sciences, Beijing 100081 ) Key Laboratory of Agricultural Ministry for Crop Genetics and Breeding Institute of Crop Breeding and Cultivation Chinese Academy of Agricultural Sciences Beijing 100081

The immature embryos of wheat plants, cv. Jing 411, 12 - 14 days after pollination, were cultured on SD2 medium for callus induction. After 10 days culture, 800 wheat calli were bombarded by biolistic particle coated with the DNA of plasmid pBI121-2 harboring both galanthus nivalis agglutinin gene and bar gene. 67 green plants were finally regenerated from the bombardment calli on selection medium containing 4mg/L Basta. The results of bioassay by both inoculating wheat aphids onto the plants and applying Basta solution of 50 mg/L and 75 mg/L onto the wheat leaves in the field, and the molecular analysis, such as PCR and Southern blotting, indicated that 8 T2 plants contaning the target genes were obtained.

关键词： galanthus nivalis agglutinin bar gene Particle bombardment Transgenic wheat

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