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| Cloning and Functional Identification of the Promoters and Terminator from Populus tomentosa Chloroplast genes |
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Abstract Abstract: Using six pairs of specific primers, DNA fragments containing the promoter of 16S rRNA gene and the promoter and terminator of psbA gene, were obtained by PCR and sequenced from Populus tomentosa chloroplast genomic DNA. Sequence analyzing results showed that the functional domains such as -35 domain, -10 domain and the translation control sequence motif in the two cloned promoters and the “TAA” sequence in the cloned terminator were also confirmed. New primers were synthetized, the promoter Prrn, PpsbA and the terminator TpsbA were cloned correctly and the prokaryotic expression vectors of GFP were constructed by using Prrn with rbcL RBS sequence, PpsbA and T7 promoter, and the terminator TpsbA, respectively. Prokaryotic expression result showed that the promoter Prrn, PpsbA and the terminator TpsbA we cloned were functional and were constitutive expressed in E.coli BL21. It slso showed that the RBS sequence was very important in gene expression.
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Received: 18 September 2006
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