Cloning and Prokaryotic Expression of O-methyltransferase from Camellia sinensis

MA Cheng-ying; SHI Jiang; LV Hai-peng; ZHANG Yue; TAN Jun-feng; GUO Li; PENG Qun-hua; LIN Zhi

doi:10.13305/j.cnki.jts.2013.06.014

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Journal of Tea Science ›› 2013, Vol. 33 ›› Issue (6) : 532-540. DOI: 10.13305/j.cnki.jts.2013.06.014

Cloning and Prokaryotic Expression of O-methyltransferase from Camellia sinensis

MA Cheng-ying, SHI Jiang, LV Hai-peng, ZHANG Yue, TAN Jun-feng, GUO Li, PENG Qun-hua, LIN Zhi^*

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Abstract

A full length cDNA of O-methyltransferase gene was obtained from Camellia sinensis and the prokaryotic expression vector for this gene was constructed. Based on total RNA from tea leaves, a O-methyltransferase cDNA sequence of tea was obtained by RT-PCR and RACE. The whole cDNA sequence 1β280βbp which contains an ORF of 1β068βbp and encodes 355 amino acids. The putative protein of this gene had an isoelectric point of 5.68 and a calculated molecular weight of 39.1βkD. The amino acid sequence of tea O-methyltransferase showed 73%, 71% identity with that of Vitis vinifera and Ricinus communis respectively. The coding sequence had been cloned into pET-28a and transformed into the host BL21. Results of SDS-PAGE showed that the specific fusion protein was successfully induced to express by IPTG.

Key words

O-methyltransferase / cloning / prokaryotic expression / tea

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MA Cheng-ying, SHI Jiang, LV Hai-peng, ZHANG Yue, TAN Jun-feng, GUO Li, PENG Qun-hua, LIN Zhi. Cloning and Prokaryotic Expression of O-methyltransferase from Camellia sinensis[J]. Journal of Tea Science. 2013, 33(6): 532-540 https://doi.org/10.13305/j.cnki.jts.2013.06.014

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