| 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].,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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