杜氏盐藻过氧化物酶1酵母双杂交诱饵质粒的构建

摘要

To construct and test whether the yeast two-hybrid bait plasmid for peroxiredoxin 1 from Dunaliella salina has self-activation and toxicity on yeast strains .Methods:The ORF of DsPrdx1 was obtained by PCR amplification , and then was inserted into the yeast expression plasmid pGBKT 7 to construct a recombinant bait plasmid pGBKT 7-Prdx1. After being identified by double digestion , the plasmid was respectively transformed into the yeast strains Y 187 and AH109 using PEG/LiAc method.Subsequently , the self-activation and toxicity were respectively examined .Results and Conclu-sion:The expression products of the recombinant bait plasmid have no self-activation or toxic effects on both Y 187 and AH109 strains,thus may be used for yeast two-hybrid system.%目的：构建杜氏盐藻过氧化物酶1（ DsPrdx1）的酵母双杂交诱饵质粒并检测其对酵母细胞有无自激活及毒害作用。方法：PCR扩增DsPrdx1的开放阅读框，将扩增出的基因片段插入酵母表达质粒pGBKT7中构建诱饵质粒。经酶切鉴定正确后，用PEG/LiAc法将构建的诱饵质粒分别转化到酵母菌株Y187和AH109中，检测诱饵蛋白对酵母菌有无自激活和毒性作用。结果与结论：构建的酵母双杂交诱饵质粒pGBKT7-Prdx1对Y187和AH109既无自激活又无毒害作用，可用于酵母双杂交系统。