IFA筛选经EMS诱导的沙眼衣原体突变株

目的:用甲基磺酸乙酯(Ethylmethylsulfone,EMS)诱导D型沙眼衣原体突变,利用间接免疫荧光法筛选出突变菌株,为研究不同衣原体基因的功能提供实验依据。方法:将D型沙眼衣原体标准株接种Mc Coy细胞,加入EMS诱导突变,收集存活菌株,利用空斑实验进行衣原体的分离和纯化,并用不同衣原体蛋白单克隆抗体做间接免疫荧光实验筛选突变株。结果:用间接免疫荧光筛选经EMS作用的沙眼衣原体,筛选出三株包涵体形态偏小的菌株(56#、58#、95#),一株圆形包涵体的突变株(61#)和一株D413N表达阴性的突变菌株(83#)。结论:用EMS作为诱导剂诱导D型沙眼衣原体突变,并成功筛选出三种突变株。为寻找衣原体功能基因与衣原体表型之间的联系奠定了实验基础。

Screening of EMS-induced Chlamydia Trachomatis Mutants by IFA

Objective:To induce Chlamydia trachomatis mutants by EMS and screen mutants by indirecting immunofluorescence(IFA), to lay experiment basis for searching Chlamydia functional genome.Methods:McCoy cell was infected with C. trachomatisserovar D and treated with EMS, the survivors harvested from EMS-treated cultures were separated and purified by plaque assay. Themutants were screened and identified by IFA with different monoclonal antibodies.Results:Three strains with small inclusion body, onestrain with round inclusion body and one strain with D413N-negative were screened out by IFA and purified by plaque assay.Conclusion:Mutant strains were successfully screened out from C. trachomatis serovar D treated with EMS, which lays an experimental basisfor further study on the relationships between functional genomes and phenotype of Chlamydia.