Construction and Expression of Fusion Protein Eukaryotic Expression Vector of Polyphenol Oxidase Gene of Tea Plant (Camellia sinensis)

WANG Nai-dong; ZHANG Li-xia; HUANG Xiao-qin; HAN Xiao-yang; LI Zhi

doi:10.13305/j.cnki.jts.2012.03.012

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Journal of Tea Science ›› 2012, Vol. 32 ›› Issue (3) : 269-275. DOI: 10.13305/j.cnki.jts.2012.03.012

Construction and Expression of Fusion Protein Eukaryotic Expression Vector of Polyphenol Oxidase Gene of Tea Plant (Camellia sinensis)

WANG Nai-dong^1,2, ZHANG Li-xia^1,2*, HUANG Xiao-qin^1,2, HAN Xiao-yang^1,2, LI Zhi^1,2

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Abstract

The polyphenol oxidase (PPO) cds sequence was amplified by PCR from the genome of Yingshuang tea plant with the high fidelity polymerase pfu. The cds sequence was recombinanted into the eukaryotic expression vector pPICZa with the designed primer, and the fusion protein eukaryotic expression vector of tea polyphenol oxidase “pPICZa-PPO” was succeeded to be constructed, and then it was transformed into Pichia pastoris GS115. Therefore, several positive converters were screened. After those positive converters were induced by methanol,the target protein was successfully detected in the culture medium by using Western-Blotting method. The detection result of enzyme activity showed that the induced products had a relatively high enzyme activity and can correctly play their biological activity.

Key words

Camellia sinensis / Pichia pastoris / fusion protein eukaryotic expression vector / inducible expression / PPO

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WANG Nai-dong, ZHANG Li-xia, HUANG Xiao-qin, HAN Xiao-yang, LI Zhi. Construction and Expression of Fusion Protein Eukaryotic Expression Vector of Polyphenol Oxidase Gene of Tea Plant (Camellia sinensis)[J]. Journal of Tea Science. 2012, 32(3): 269-275 https://doi.org/10.13305/j.cnki.jts.2012.03.012

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