Spodoptera LituraTolerant Transgenic Tobacco Plants Expressing Genetically ModifiedCRY 1ABGenes fromBacillus Thuringiensis

Christov, N.; Shimizu, N; Ohkawa, Hideo

doi:10.1080/13102818.1998.10818957

Cited by 3 publications

(1 citation statement)

References 21 publications

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“…Some of the specific factors include premature polyadenylation, mRNA instability, improper codon usage, aberrant splicing, and transgene silencing (Diehn et al 1996;Christov et al 1998;Grec et al 2000;Ibrahim et al 2001;Ayliffe et al 2004;Geyer et al 2007Geyer et al , 2010Lin et al 2009). A/T richness of the coding region has been identified as a general signal for RNA processing in plants (Luehrsen et al 1994;Luehrsen and Walbot 1994a, b;Baynton et al 1996;Misztal et al 2004;Narsai et al 2007).…”

Section: Introductionmentioning

confidence: 98%

Spurious polyadenylation of Norovirus Narita 104 capsid protein mRNA in transgenic plants

et al. 2011

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Noroviruses are members of the family Caliciviridae, and cause a highly communicable gastroenteritis in humans. We explored the potential to develop a plant-based vaccine against Narita 104 virus, a Genogroup II Norovirus. In stably transgenic potato, we obtained very poor expression of Narita 104 virus capsid protein (NaVCP) despite the use of a strong constitutive promoter (dual enhancer 35S) driving the native coding sequence. We identified potentially detrimental sequence motifs that could mediate aberrant mRNA processing via spurious polyadenylation signals. Northern blots and RT-PCR analysis of total RNA revealed truncated transcripts that suggested premature polyadenylation. Site-directed mutagenesis to remove one potential polyadenylation near-upstream element resulted in an increased expression of NaVCP when transiently expressed in leaves of Nicotiana benthamiana. Further, cloning of the truncated cDNAs from transgenic NaVCP potato plants and transiently transfected N. benthamiana allowed us to identify at least ten different truncated transcripts resulting from premature polyadenylation of full length NaVCP transcripts. Comparative studies using real time PCR analysis from cDNA samples revealed lower accumulation of full length transcripts of NaVCP as compared to those from a gene encoding Norwalk Virus capsid protein (a related Genogroup I Norovirus) in transiently transfected plants. These findings provide evidence for impaired expression of NaVCP in transgenic plants mediated by spurious polyadenylation signals, and demonstrate the need to scrupulously search for potential polyadenylation signals in order to improve transgene expression in plants.

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