@article{lu_sivamani_azhakanandam_samadder_li_qu_2008, title={Gene expression enhancement mediated by the 5 ' UTR intron of the rice rubi3 gene varied remarkably among tissues in transgenic rice plants}, volume={279}, ISSN={["1617-4615"]}, DOI={10.1007/s00438-008-0333-6}, abstractNote={Introns are important sequence elements that modulate the expression of genes. Using the GUS reporter gene driven by the promoter of the rice (Oryza sativa L.) polyubiquitin rubi3 gene, we investigated the effects of the 5' UTR intron of the rubi3 gene and the 5' terminal 27 bp of the rubi3 coding sequence on gene expression in stably transformed rice plants. While the intron enhanced GUS gene expression, the 27-bp fused to the GUS coding sequence further augmented GUS expression level, with both varying among different tissues. The intron elevated GUS gene expression mainly at mRNA accumulation level, but also stimulated enhancement at translational level. The enhancement on mRNA accumulation, as determined by realtime quantitative RT-PCR, varied remarkably with tissue type. The augmentation by the intron at translational level also differed by tissue type, but to a lesser extent. On the other hand, the 27-bp fusion further boosted GUS protein yield without affecting mRNA accumulation level, indicating stimulation at translation level, which was also affected by tissue type. The research revealed substantial variation in the magnitudes of intron-mediated enhancement of gene expression (IME) among tissues in rice plants and the importance of using transgenic plants for IME studies.}, number={6}, journal={MOLECULAR GENETICS AND GENOMICS}, author={Lu, Jianli and Sivamani, Elumalai and Azhakanandam, Kasi and Samadder, Partha and Li, Xianggan and Qu, Rongda}, year={2008}, month={Jun}, pages={563–572} }
 @article{azhakanandam_weissinger_nicholson_qu_weissinger_2007, title={Amplicon-plus Targeting Technology (APTT) for rapid production of a highly unstable vaccine protein in tobacco plants (vol 64, pg 619, 2007)}, volume={64}, ISSN={["0167-4412"]}, DOI={10.1007/s11103-007-9168-5}, number={5}, journal={PLANT MOLECULAR BIOLOGY}, author={Azhakanandam, Kasi and Weissinger, Sandra M. and Nicholson, Jennifer S. and Qu, Rongda and Weissinger, Arthur K.}, year={2007}, month={Jul}, pages={619–619} }
 @article{azhakanandam_weissinger_nicholson_qu_weissinger_2007, title={Amplicon-plus targeting technology (APTT) for rapid production of a highly unstable vaccine protein in tobacco plants}, volume={63}, DOI={10.1007/s11103-006-9096-9}, abstractNote={High-level expression of transgenes is essential for cost-effective production of valuable pharmaceutical proteins in plants. However, transgenic proteins often accumulate in plants at low levels. Low levels of protein accumulation can be caused by many factors including post-transcriptional gene silencing (PTGS) and/or rapid turnover of the transgenic proteins. We have developed an Amplicon-plus Targeting Technology (APTT), by using novel combination of known techniques that appears to overcome both of these factors. By using this technology, we have successfully expressed the highly-labile L1 protein of canine oral papillomavirus (COPV L1) by infecting transgenic tobacco plants expressing a suppressor of post-transcriptional gene silencing (PTGS) with a PVX amplicon carrying a gene encoding L1, and targeting the vaccine protein into the chloroplasts. Further, a scalable "wound-and-agrospray" inoculation method has been developed that will permit high-throughput Agrobacterium inoculation of Nicotiana tabacum, and a spray-only method (named "agrospray") for use with N. benthamiana to allow large-scale application of this technology. The good yield and short interval from inoculation to harvest characteristic of APTT, combined with the potential for high-throughput achieved by use of the agrospray inoculation protocol, make this system a very promising technology for producing high value recombinant proteins, especially those known to be highly labile, in plants for a wide range of applications including producing vaccines against rapidly evolving pathogens and for the rapid response needed to meet bio-defense emergencies.}, number={3}, journal={Plant Molecular Biology}, author={Azhakanandam, K. and Weissinger, S. M. and Nicholson, J. S. and Qu, R. D. and Weissinger, A. K.}, year={2007}, pages={393–404} }