Construction of an Expression Vector for Production and Purification of Human Somatostatin in Escherichia coli |
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Authors: | Sergi Maicas Ismaïl Moukadiri Almudena Nieto Eulogio Valentín |
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Affiliation: | 1. Department of Plant Biotechnology, Centre for Plant Molecular Biology and Biotechnology, Tamil Nadu Agricultural University, Coimbatore, 641003, Tamil Nadu, India 2. Centre for Plant Protection Studies, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India 3. Horticulture College and Research Institute, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India 4. Tapioca and Castor Research Station, Tamil Nadu Agricultural University, Yethapur, Salem, Tamil Nadu, India 5. Central Tuber Crops Research Institute, Trivandrum, Kerala, India
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Abstract: | Cassava mosaic disease, caused by cassava mosaic geminiviruses are transmitted by Bemisia tabaci. The B. tabaci adults from colonies reared on virus free cassava plant produced from apical meristem culture was studied to determine their ability to transmit Indian cassava mosaic virus (ICMV) and Sri Lankan cassava mosaic virus (SLCMV) from cassava to cassava. Virus free plants were confirmed by polymerase chain reaction (PCR) using geminivirus degenerate primers. The virus acquisition access period (AAP) of 48 h on virus infected cassava leaves and 48 h virus inoculation access periods on virus free healthy leaves were investigated. Both ICMV and SLCMV were absolutely transmitted by whiteflies reared on cassava. Virus specific primers were designed in the replicase region and used to detect virus in B. tabaci after different AAP. The PCR amplified replicase genes from virus transmitted cassava leaves were cloned the plasmid DNA was isolated from a recombinant colony of E. coli DH5α after their confirmation by colony PCR and sequenced them. The nucleotide sequences obtained from automated DNA sequencing were confirmed as ICMV and SLCMV replicase gene after homology searching by BLAST and found to be a new isolates. The nucleotide sequences of new isolates were submitted in GenBank (accession number JN652126 and JN595785). |
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