摘要
通过RACE技术克隆到盐藻(DunaliellasalinaTeod.)果糖 1,6 二磷酸醛缩酶(DsALDP)的全长cDNA.对其序列分析及在GeneBank中进行同源搜索,发现DsALDP基因属于果糖 1,6 二磷酸醛缩酶基因家族,其与植物叶绿体果糖 1,6 二磷酸醛缩酶(AldP)基因的一致性为73%~66%.Northern杂交结果及醛缩酶活性分析均表明它确为在盐诱导下特异表达.将DsALDPcDNA构建到pET32a载体上,转入E.coliBL21进行表达分析.IPTG诱导后可以检测到一62ku基因产物大量表达.经不同浓度NaCl处理,表达DsALDP基因产物的细菌耐盐性明显高于对照组.
The fulllength cDNA of fructose1, 6diphosphate aldolase in Dunaliella salina was cloned by rapid amplification of cDNA ends (RACE). It was shown from sequence analysis and BlastP search in GeneBank that DsALDP belonged to fructose1, 6diphosphate aldolase family, and the identities of DsALDP to AldP of other plants were 73%66%. It was confirmed by northern blot and aldolase activity analysis that DsALDP was specially expressed under NaCl stress. After insertion of DsALDP to vector pET32a, the recombined plasmid was transferred into E. coli BL21 to analyze its expression. After adding IPTG, a Large amount of 62 ku fusion gene product was detected. The bacteria were cultured in media with different NaCl concentration. As a result, the bacteria expressing DsALDP exhibited a higher salt tolerance with increasing NaCl concentration than bacteria with no DsALDP expression.
出处
《复旦学报(自然科学版)》
CAS
CSCD
北大核心
2002年第5期593-595,共3页
Journal of Fudan University:Natural Science
基金
上海市教委曙光计划资助项目(98SG47)