New gene construction strategy in T-DNA vector to enhance expression level of sweet potato sporamin and insect resistance in transgenic Brassica oleracea

Huai-Ju Chen; Shu-Jen Wang; Chien-Cheng Chen; Kai-Wun Yeh

doi:10.1016/j.plantsci.2006.04.003

New gene construction strategy in T-DNA vector to enhance expression level of sweet potato sporamin and insect resistance in transgenic Brassica oleracea

Plant Sci. 2006 Sep;171(3):367-74. doi: 10.1016/j.plantsci.2006.04.003. Epub 2006 May 5.

Authors

Huai-Ju Chen¹, Shu-Jen Wang, Chien-Cheng Chen, Kai-Wun Yeh

Affiliation

¹ Graduate Institute of Plant Biology, College of Life Science, National Taiwan University, Taipei 106, Taiwan.

PMID: 22980206
DOI: 10.1016/j.plantsci.2006.04.003

Abstract

Sporamin, an abundant storage protein in tuberous roots of sweet potato, possesses strong inhibitory activity against trypsin and pest-resistance. To promote consistent high-level expression of sporamin and insect resistance in transgenic Brassica plants, a wound-responsive sporamin promoter (Pspoa) alone or combined with matrix-attached-region-like DNA segment (spoMAR) were constructed for driving sporamin cDNA. The results showed the transgenic plants containing Pspoa-drived sporamin and spoMAR displayed the highest level and low inter-transformant variability of sporamin expression, and the ability of insect resistance of transformants positively correlated with sporamin activity. Furthermore, expressions of Pspoa-drived sporamin especially combined with the spoaMAR retains high and steady levels in the T(1) and T(2) generations, in marked contrast to the variable expression patterns observed in CaMV35S promoter-driven transformants. This study evidently indicates that the Pspoa and spoaMAR would be very efficient for high transgene expression in plants and obtaining inherently stable transformants in consecutive progenies.