Development of an intra-molecularly shuffled efficient chimeric plant promoter from plant infecting Mirabilis mosaic virus promoter sequence

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• 作者：Sefali Acharya ; Soumika Sengupta ; Sunita Patro ; Sukumar Purohit ; Sabindra K. Samal ; Indu B. Maiti ; Nrisingha Dey
• 关键词：Chimeric promoter ; IL-24 ; MMV ; GFP ; GUS
• 刊名：Journal of Biotechnology
• 出版年：10 January, 2014
• 年：2014
• 卷：169
• 期：Complete
• 页码：103-111
• 全文大小：1737 K

文摘

We developed an efficient chimeric promoter, MUASMSCP, with enhanced activity and salicylic acid (SA)/abscisic acid (ABA) inducibility, incorporating the upstream activation sequence (UAS) of Mirabilis mosaic virus full-length transcript (MUAS, 鈭?97 to 鈭?8) to the 5鈥?end of Mirabilis mosaic virus sub-genomic transcript (MSCP, 鈭?06 to 鈭?25) promoter-fragment containing the TATA element. We compared the transient activity of the MUASMSCP promoter in tobacco/Arabidopsis protoplasts and in whole plant (Petunia hybrida) with the same that obtained from CaMV35S and MUAS35SCP promoters individually. The MUASMSCP promoter showed 1.1 and 1.5 times stronger GUS-activities over that obtained from MUAS35SCP and CaMV35S promoters respectively, in tobacco (Xanthi Brad) protoplasts. In transgenic tobacco (Nicotiana tabacum, var. Samsun NN), the MUASMSCP promoter showed 1.1 and 2.2 times stronger activities than MUAS35SCP and CaMV35S² promoters respectively. We observed a fair correlation between MUASMSCP-, MUAS35SCP- and CaMV35S²-driven GUS activities with the corresponding uidA-mRNA level in transgenic plants. X-gluc staining of transgenic germinating seed-sections and whole seedlings also support above findings. Protein-extracts made from tobacco protoplasts expressing GFP and human-IL-24 genes driven individually by the MUASMSCP promoter showed enhanced expression of the reporters compared to that obtained from the CaMV35S promoter. Furthermore, MUASMSCP-driven protoplast-derived human IL-24 showed enhanced cell inhibitory activity in DU-145 prostate cancer cells compared to that obtained from the CaMV35S promoter. We propose chimeric MUASMSCP promoter developed in the study could be useful for strong constitutive expression of transgenes in both plant/animal cells and it may become an efficient substitute for CaMV35S/CaMV35S² promoter.