田颖新 杨长生 贾天军 王冰 杨瑞宁 邱红.IFA筛选经EMS诱导的沙眼衣原体突变株[J].现代生物医学进展英文版,2016,16(5):852-856. |
IFA筛选经EMS诱导的沙眼衣原体突变株 |
Screening of EMS-induced Chlamydia Trachomatis Mutants by IFA |
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DOI: |
中文关键词: EMS 突变菌株 空斑实验 间接免疫荧光 |
英文关键词: EMS Mutant strain Plaque Assay IFA |
基金项目:南京军区面上项目(12MA038) |
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中文摘要: |
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McCoy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。 |
英文摘要: |
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. trachomatis
serovar D and treated with EMS, the survivors harvested from EMS-treated cultures were separated and purified by plaque assay. The
mutants were screened and identified by IFA with different monoclonal antibodies.Results:Three strains with small inclusion body, one
strain 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 basis
for further study on the relationships between functional genomes and phenotype of Chlamydia. |
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