Development of an intra-molecularly shuffled efficient chimeric plant promoter from plant infecting Mirabilis mosaic virus promoter sequence.
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, -297 to -38) to the 5' end of Mirabilis mosaic virus sub-genomic transcript (MSCP, -306 to -125) 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 CaMV35S(2) promoters respectively. We observed a fair correlation between MUASMSCP-, MUAS35SCP- and CaMV35S(2)-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/CaMV35S(2) promoter.
Division of Gene Function and Regulation, Institute of Life Sciences, Department of Biotechnology, Government of India, Nalco Square, Chandrasekharpur, Bhubaneswar 751 023, Odisha, India. Electronic address: sefaliacharya@gmail.com.
Sefali
S
Acharya
Division of Gene Function and Regulation, Institute of Life Sciences, Department of Biotechnology, Government of India, Nalco Square, Chandrasekharpur, Bhubaneswar 751 023, Odisha, India. Electronic address: soumika.sengupta@gmail.com.
Soumika
S
Sengupta
Division of Gene Function and Regulation, Institute of Life Sciences, Department of Biotechnology, Government of India, Nalco Square, Chandrasekharpur, Bhubaneswar 751 023, Odisha, India. Electronic address: sunitapatro07@gmail.com.
Sunita
S
Patro
Division of Gene Function and Regulation, Institute of Life Sciences, Department of Biotechnology, Government of India, Nalco Square, Chandrasekharpur, Bhubaneswar 751 023, Odisha, India. Electronic address: sukumarpurohit1988@gmail.com.
Sukumar
S
Purohit
Division of Gene Function and Regulation, Institute of Life Sciences, Department of Biotechnology, Government of India, Nalco Square, Chandrasekharpur, Bhubaneswar 751 023, Odisha, India. Electronic address: sabindra.kr.samal@gmail.com.
Sabindra K
SK
Samal
Plant Genetic Engineering & Service, KTRDC, University of Kentucky, Lexington, KY 40546-0236, USA.
Indu B
IB
Maiti
Division of Gene Function and Regulation, Institute of Life Sciences, Department of Biotechnology, Government of India, Nalco Square, Chandrasekharpur, Bhubaneswar 751 023, Odisha, India. Electronic address: dey@ils.res.in.
Nrisingha
N
Dey
D000040:Abscisic Acid / Q000494:pharmacology; D000818:Animals; D017360:Arabidopsis / Q000235:genetics; D017796:Caulimovirus / Q000235:genetics*; D045744:Cell Line, Tumor; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D006801:Humans; D032306:Petunia / Q000235:genetics; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011523:Protoplasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics; D020156:Salicylic Acid / Q000494:pharmacology; D014026:Nicotiana / Q000235:genetics
D011993:Recombinant Fusion Proteins; D000040:Abscisic Acid; D020156:Salicylic Acid
Chimeric promoter; GFP; GUS; IL-24; MMV