Functional analysis of the 3' control region of the potato wound-inducible proteinase inhibitor II gene.

Abstract

Proteinase inhibitor genes are expressed strongly in specific plant tissues under both developmental and environmental regulation. We have studied the role of the 3' control region of the potato proteinase inhibitor II gene (PI-II) that is inducible in leaves in response to herbivore attacks or other severe wounding. Comparison of the terminator from the PI-II gene with two different terminators from the 6b and 7 genes, driven by a common PI-II promoter-cat fusion molecule, indicated that the PI-II terminator provided the most efficient expression of cat. The PI-II terminator also caused a significantly elevated cat gene expression driven by the cauliflower mosaic virus 35S promoter. The increase in the level of expression is probably not due to the presence of an enhancer element in the PI-II terminator region, but to cis-acting elements involved in mRNA processing or stability. Both transient and stable transformation analyses of the deletion mutants in the 3'-flanking sequence indicated that about a 100-base pair DNA fragment surrounding the polyadenylation site is essential for the efficient gene expression. This region seems to consist of several regulatory elements, including the conserved sequence, CGTGTCTT, which is located 9 bases downstream from the polyadenylation site. The elements appear to contribute to the increased stability of mRNAs containing the PI-II terminator.

Specific Tandem Repeats Are Sufficient for Paramutation-Induced Trans-Generational Silencing

“…GUS is the beta-glucuronidase gene from E. coli [29]. PinII is the 3′ untranslated region from the potato proteinaseII gene [61]. The p35S::GUS construct, in which GUS expression is driven by the minimal −90 35S promoter, was used as a control.…”

Bioencapsulation of the hepatitis B surface antigen and its use as an effective oral immunogen

“…HBE contained the 1.4kb globulin1 promoter (NCBI Gene ID 732801) and expressed a BAASS:HBsAg fusion protein. All constructs contained a potato protease inhibitor II ( Pin II ) termination sequence [37] and the glyphosate resistance gene [38]. …”

Commercial Plant-Produced Recombinant Avidin

“…All movement between cloning vectors was done with restriction enzyme digestion and ligation. The expression cassette with the constitutive maize ubiquitin promoter (Christensen et al 1992) and the pinII terminator (An et al 1989) was built separately from the herbicide selection vector for co-bombardment of maize callus tissue. Selection was on the herbicide, bialaphos, using the bar gene (White et al 1990) driven by the CaMV 35S promoter.…”

“…This observation was later followed up and shown to be very effective to prevent postharvest insect damage while not interfering with metabolism in mammals (Kramer et al 2000). The transgenic maize plants had a secondary phenotype in that they could confer male sterility and have been suggested as a containment mechanism for transgenic traits in the field (Albertsen et al 1999).…”