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一种嵌合的花椰菜花叶病毒35S苏云金芽孢杆菌杀虫蛋白基因在转基因烟草中的表达

Expression of a chimeric CaMV 35S Bacillus thuringiensis insecticidal protein gene in transgenic tobacco.

作者信息

Carozzi N B, Warren G W, Desai N, Jayne S M, Lotstein R, Rice D A, Evola S, Koziel M G

机构信息

CIBA-Geigy Agricultural Biotechnology Research Unit, Research Triangle Park, NC 27709.

出版信息

Plant Mol Biol. 1992 Nov;20(3):539-48. doi: 10.1007/BF00040612.

Abstract

Insecticidal transgenic tobacco plants containing a truncated Bacillus thuringiensis cryIA(b) crystal protein (ICP) gene expressed from the CaMV 35S promoter were analyzed for ICP gene expression under field and greenhouse conditions over the course of a growing season. We present new information on temporal and tissue-specific expression of a CaMV 35S/cryIA(b) gene. Levels of cryIA(b) protein and mRNA were compared in both homozygous and hemizygous lines throughout plant development. Levels of ICP mRNA and protein increased during plant development with a pronounced rise in expression at the time of flowering. Homozygous ICP lines produced higher levels of ICP than the corresponding hemizygous lines. ELISA analysis of different tissues in the tobacco plant showed ICP gene expression in most tissues with a predominance of ICP in older tissue. All transgenic ICP tobacco lines which were studied in the field and greenhouse contained 400 ng to 1 microgram ICP per gram fresh weight in leaves from the mid-section of the plant at flowering. The amounts of ICP produced by field lines were directly comparable to levels observed in greenhouse-grown plants.

摘要

对含有截短的苏云金芽孢杆菌cryIA(b)晶体蛋白(ICP)基因的杀虫转基因烟草植株进行了分析,该基因由花椰菜花叶病毒(CaMV)35S启动子表达,在一个生长季节的田间和温室条件下检测其ICP基因表达情况。我们提供了关于CaMV 35S/cryIA(b)基因的时间和组织特异性表达的新信息。在整个植株发育过程中,对纯合和半合子株系中的cryIA(b)蛋白和mRNA水平进行了比较。ICP mRNA和蛋白水平在植株发育过程中升高,在开花时表达明显增加。纯合ICP株系产生的ICP水平高于相应的半合子株系。对烟草植株不同组织的ELISA分析表明,大多数组织中都有ICP基因表达,且老龄组织中ICP占主导。在田间和温室中研究的所有转基因ICP烟草株系,在开花期植株中部叶片中每克鲜重含有400纳克至1微克ICP。田间株系产生的ICP量与温室种植植株中观察到的水平直接可比。

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