铜绿假单胞菌外毒素A全长基因的扩增与克隆

Amplification and cloning of exotoxin A gene from Pseudomonas aeruginosa

目的 从绿脓杆菌中提取染色体DNA ,PCR扩增及克隆外毒素A全长基因 ,为实现外毒素A的高效表达奠定基础。方法 从绿脓杆菌培养物中提取染色体DNA ,PCR扩增外毒素A全长基因 ,A -T克隆入本室自行构建的pSK -T载体中 ,对重组质粒进行酶切与测序鉴定。结果与结论 成功地从高GC含量的绿脓杆菌染色体DNA中扩增到长片段的外毒素A全基因并进行了克隆与鉴定。

Aim To amplify and clone the full length exotoxin A gene Methods Extract genomic DNA from P aeruginosa culture By using PCR technique,exotoxin A gene was amplified and linked to pSK T vector,then transformed to E coli DH10B The recombinant plasmid was identified by BamHⅠ and BglⅡ digestion The recombinant plasmid was also identified by sequencing Result and Conclusion Successfully amplified long exotoxin A gene from GC rich P aeruginosa genomic DNA The exotoxin A gene has cloned and identified