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| Cloning and Analysis of the RNA Polymerase i-Promoter of Nicotiana benthaminana |
| LI Zhi-ying1,2, MU Hong-zhen2, GAO Ding-mei2, DING Guo-ping2, MA Ting2, WANG Sheng1,2 |
1. Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resources in the Western China, Yinchuan 750021, China;
2. School of Life Science, Ningxia University, Yinchuan 750021, China |
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Abstract Promoter is an important transcriptional regulatory element, which controls the levels and patterns of gene expression. RNA Pol Ⅰ is a cellular enzyme that is abundantly expressed in cells and transcribes rRNA precursor lacking a 5' cap, a 3' poly (A) tail and introns. Thus, viral RNA synthesized in cells transfected with Pol Ⅰ-driven plasmids containing viral genomic cDNA has precise sequences. This may increase the expression level and biological safety of the plant RNA-based expression system. In addition, Nicotiana benthaminana is a kind of model organism which widely used in plant bioreactor and plant pathology. However, there is no nucleic acid sequence data about its RNA Pol Ⅰ promoter for now. Therefore, Nicotiana benthaminana Pol Ⅰ promoter sequence we cloned and analysized its transcription initiation site (TIS). The 514 bp RNA pol Ⅰ promoter sequence (KC352713) was amplified by semi-net PCR form Nicotiana benthaminana and the third A residue in the core sequence TATA (G) TA (N) GGGGG was predicted to be the transcription initiation site (TIS) by comparison of sequences flanking RNA pol Ⅰ promoter TIS in silicon. The predicted result was conformed by the 5' RACE and sequence analysis. This result will be beneficial for the further research of both Pol Ⅰ promoter and Pol Ⅰ-based plant virus expression vectors.
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Received: 21 October 2013
Published: 25 January 2014
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