Analysis of unstable RNA transcripts of insecticidal crystal protein genes of Bacillus thuringiensis in transgenic plants and electroporated protoplasts.
We have examined expression of several insecticidal crystal protein (ICP) genes of Bacillus thuringiensis in transgenic tobacco plants and electroporated carrot protoplasts. We determined that low levels of lepidopteran toxin cryIA(b) ICP gene expression in plants and electroporated carrot cells is due to RNA instability. We used a series of 3' deleted by cryIA(b) constructs directed by the cauliflower mosaic virus 35S promoter to demonstrate that this instability is minimally contained in the first 579 bases of the gene in both systems. This instability may result from 5'----3' as well as 3'----5' RNA metabolism. The coleopteran toxic cryIIIA gene was also examined in electroporated carrot cells, and found to be poorly expressed. A model for improvement of ICP RNA stability in plants is presented.
Murray
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EE
Agrigenetics Advanced Research Corporation, Madison, WI 53716.
Rocheleau
T
T
Eberle
M
M
Stock
C
C
Sekar
V
V
Adang
M
M
D001413:Bacillus thuringiensis / Q000235:genetics*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins; D015152:Blotting, Northern; D002460:Cell Line; D002872:Chromosome Deletion; D003062:Codon / Q000235:genetics; D004731:Endotoxins; D015870:Gene Expression / Q000502:physiology; D006460:Hemolysin Proteins; D009029:Mosaic Viruses / Q000235:genetics; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic / Q000235:genetics; D011523:Protoplasts / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D014026:Nicotiana / Q000378:metabolism; D014170:Transformation, Genetic / Q000502:physiology
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D003062:Codon; D004731:Endotoxins; D006460:Hemolysin Proteins; D012333:RNA, Messenger; C030956:insecticidal crystal protein, Bacillus Thuringiensis