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《Journal of Huazhong Agricultural University》 2009-01
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PCR Amplication,Prokaryote Expression of SHV-12 Type Extend-spectrum β-lactamase Produced by Shigella Isolated from Fowl and Its Related Characteristics

SUN Ya-wei HU Gong-zheng CHEN Hong-ying LIU Jian-huaYUAN Li DENG Li-xin YUAN Ye-you(College of Animal Husbandary and Veterinary Medicine Engerning,Henan Agricultural University,Zhengzhou 450002,China)  
SHV-12 type extended-spectrum β-lactamase genes produced by 5 shigella isolates from fowl was cloned and expressed under optimized condition and related characteristics of the expressed proteins were studied.The results showed that the taget fragment was obtained after the constructed recombinant plasmids was digested by EcoRⅠand Xhol Ⅰ.The constructed recombinant plasmids expressed recombinant proteins which possessed responding ESBLs activities.The optimized conditions of expressing recombinant strains S-BL21 was:IPTG concenration was 0.5 mmol/L,the induced temperature was 37℃,the induced time was 5 h.The SHV-12 type ESBLs has power affinity to ceftifur sodium with Km value of 1.29 μmol and has more difficulty to hydrolyse ceftriaxone with Km value of 24.13 μmol.The inhibition rate of tazobactam and sulbactam on hydrolyzing ceftriaxone and ceftiofur of SHV-12 type ESBLs was 21.01%,25.51%,34.99%,and 35.49% respectively.
【Fund】: 国家自然科学基金项目(30471307);; 河南农业大学创新基金项目(2007)资助
【CateGory Index】: S852.61
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