萝卜过氧化物酶基因RsPrx1在毕赤酵母中的表达及其抗性

Expression of Radish Peroxidase Gene RsPrx1 in Pichia pastoris and Studies on Its Tolerance

从成熟心里美萝卜肉质根的皮中提取总RNA,采用RT-PCR方法获得过氧化物酶基因RsPrx1的cDNA,将其克隆到pGEM-T载体中,并构建毕赤酵母重组表达载体pPIC9KH-RsPrx1。用L iC l法转化野生型毕赤酵母GS115获得重组转化子。用不同浓度的NaC l和H2O2处理重组毕赤酵母转化子,结果表明该转化子具有一定的抗盐和抗氧化能力。建立了RsPrx1重组毕赤酵母表达体系,能够有效表达具有生物活性的外源过氧化物酶。

RsPrxl cDNA was amplified by reverse transcription-polymerase chain reaction （RT - PCR） from the peel of Chinese red radish （Raphanus sativus L. ） root. The full-length of RsPrx1 cDNA was sequenced. Then it was sub-cloned into Pichia pastoris expression plasmid pPIC9 KH. RsPrx1 gene was integrated into the genome of Pichia pastoris and successfully expressed by inducing. The recombinants could show certain resistance to NaCl and H2O2. An efficient expression system of Pichia pastoris was constructed, which was the base of large scale production and functional analysis.