论文部分内容阅读
目的 构建白喉杆菌染色体整合质粒 ,建立使外源基因置换白喉毒素基因的方法。方法 PCR方法 ,寡核苷酸合成方法及其他分子克隆技术。结果 使用以上方法准确制备了所需特定碱基序列位置的 2个与白喉毒素基因相关的染色体同源片段 ,插入pG +host5后又引入转录终止子和抗生素抗性基因 ,从而产生具有可接入外源基因并与白喉杆菌染色体进行双交换同源重组能力的质粒pLB2 3。用此质粒对白喉杆菌PW8进行了染色体整合 ,获得了其遗传变异株。结论 所构建的整合质粒能够用于白喉杆菌染色体的同源重组 ,实验为将外源基因引入白喉杆菌染色体进行表达奠定了基础。
Objective To construct the chromosomal integration plasmid of diphtheria diphtheriae, and to establish a method for exogenous gene replacement of diphtheria toxin gene. Methods PCR methods, oligonucleotide synthesis methods and other molecular cloning techniques. Results Two chromosomal homology fragments related to diphtheria toxin gene were precisely prepared by the above method. After inserted into pG + host5, transcriptional terminator and antibiotic resistance gene were introduced, Plasmid pLB2 3 that has exogenous genes and is capable of double crossover homologous recombination with the diphtheria bacillus chromosome. Chromosome integration of diphtheriae PW8 was carried out using this plasmid, and the genetic variation of the strain was obtained. Conclusion The constructed plasmid can be used for homologous recombination of Chryseobacterium diphtheriae. The experiment laid the foundation for the introduction of exogenous gene into Chryseobacterium.