高温促进含第1类整合子宿主菌的重组效率  

Increased recombination frequency catalyzed by host strains containing class 1 integrons under high temperature stress

作　　者：胡敏[1] 赵克斌[1] 杨泽华[1] 武建才[1] 

机构地区：[1]山西医科大学第一医院检验科,太原030001

出　　处：《中国药物与临床》2013年第10期1259-1262,共4页Chinese Remedies & Clinics

基　　金：山西省自然科学基金(2012011041-2)

摘　　要：目的探讨高温对大肠杆菌BL21(DE3)宿主中第1类整合子重组效率的影响。方法用pUC19为载体构建高表达整合酶的重组质粒pUCINT,随后在pACYC184载体的不同位置分别插入整合子以及氨基糖苷腺苷转移酶(aadA2)基因盒,该重组质粒命名为pACINAD,将这2种相容的重组质粒转化大肠杆菌BL21(DE3),将20株此菌株随机分成2组,每组各10株,分别在41℃和37℃培养24 h后用荧光定量聚合酶链反应(PCR)测定发生重组反应的整合子的拷贝数和总的整合子的拷贝数,前者与后者的比值即为重组效率。结果 41℃大肠杆菌BL21(DE3)宿主中所测得的重组效率为(1.8±0.5)×10-3,在37℃的重组效率为(1.8±0.5)×10-4。41℃组明显高于37℃组,差异具有统计学意义(t=9.266,P<0.01)。结论高温可显著提高由整合酶介导的aadA2基因盒的重组效率。Objective To explore the effects of high temperature stress on recombination frequency of type 1 integrons in Escherichia coli（E.coli） BL21 （DE3） hosts. Methods The recombinant plasmid pUCINT with a high yield of integrase was constructed with the pUC19 vector. This was followed by integration of the integrons and amino- glycyl adenosine transferrase （aadA 2） gene box at different locations into the pACYC 184 vector, thus forming a recom- binant vector pACINAD. The two compatible recombinant plasmids were used to transfect the E.coli BL21 （DE3） hosts. A total of 20 strains of E. coli were randomly assigned to two groups for incubation at 41℃ and 37℃for 24 hours, respectively. The copy numbers of cointegrates and total transconjugants were detected by using fluorescent polymerase chain reaction, andthe recombination frequency was expressed as the ratio of cointegrates to total transconjugants. Results The recombination frequency in host BL21 （DE3） was （1.8±0.5） ×10.3 at 41 ℃ and the re- combination frequency of aadA2 gene cassettes was （1.8±0.5）×104 at 37 ℃（t=9.266, P〈0.01）. Conclusion High temperature stress can markedly promote the integrase-mediated recombination efficiency of aadA 2 gene box.

关 键 词：整合子类 抗药性 重组效率 高温 

分 类 号：R378[医药卫生—病原生物学]