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草业科学  2014 

马铃薯软腐病菌Erwiniacarotovorasubsp.carotovora714种胞外酶基因的克隆和原核表达分析

, PP. 561-574

Keywords: 马铃薯,软腐病,致病因子,克隆,原核表达

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Abstract:

?以马铃薯软腐病菌Erwiniacarotovorasubsp.carotovora71(Ecc71)为材料,利用同源克隆技术克隆软腐病果胶酸盐裂解酶(pel)、多聚半乳糖醛酸酶(peh)、纤维素酶(cel)和蛋白酶(prt)基因,对其进行生物信息学分析,并将其克隆到原核表达载体pET28a中,将重组载体pET28a-pel、pET28a-peh、pET28a-cel和pET28a-prt转化至大肠杆菌E.coliBL21(DE3),经IPTG诱导后,SDS-PAGE电泳鉴定和酶活测定,得到了与理论推算的pel、peh、cel和prt基因表达产物分子质量相符的特异条带,并且其酶活性分别为1.672U·mL-1·h-1和0.024、112.7、16.95U·mL-1·min-1。这些结果为今后探究这4种酶之间的相互作用及马铃薯抗病性研究奠定了基础。

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