Cloning of strawberry FaEtr2 gene and its plant expression vector construction for antisense RNA

Abstract

An ethylene receptor FaEtr2 gene was amplified by Polymerase Chain Reaction (PCR) from ripening strawberry fruit. A 1049-bp PCR product (All Star-Etr2) was cloned. Sequence analysis showed that the All Star-Etr2 nucleotide sequence had 100% identity with Chandler-Etr2 from the GenBank. A pair of primers containing restriction enzyme sites were designed and used to amplify the sequenced plasmid. The PCR product was digested by the corresponding restricted enzymes and inserted between the CaMV 35S promoter and NOS terminator of expression vector pBI121 directionally. The constructed expression vector was transformed into Agrobacterium fumefeciens LBA4404 in the follow-up research to silence a ripening-related ethylene receptor FaEtr2 gene in strawberry fruits.

Keywords

strawberry / ethylene receptor / FaEtr2 gene / plant expression vector

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This research was supported by National Natural Science Foundation of China (Grant No. 30500349) and Beijing Natural Science Foundation, China (No. 6062020)

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2014 Higher Education Press and Springer-Verlag Berlin Heidelberg