10种预测的鹦鹉热衣原体Ⅲ型分泌系统效应蛋白的定位分析

doi:10.11843/j.issn.0366-6964.2020.018

Abstract

Abstract: This study aimed to investigate the intracellular localization of 10 putative effector proteins via type Ⅲ secretion system (T₃SS) in Chlamydia psittaci, laying the basis for the next structure and function analysis. In this study, computer-aided bioinformatics methods such as Effective T3 and BPBAac were used to select the possible T₃SS effector proteins of C psittaci. The recombinant plasmid pGEX -6P-1-target genes were constructed and transformed into E. coli BL21, and the recombinant proteins were induced and purified. Then BALB/c mice were immunized with the recombinant proteins to produce polyclonal antibodies, and the titre of antibodies were detected by ELISA. Indirect immunofluorescence assay (IFA) was used to detect the subcellular localization of these proteins. In our study, fourteen possible Cps T₃SS effector protein-coding genes were predicted:e.g. CPSIT_0357, CPSIT_0429, CPSIT_0461, CPSIT_0463, CPSIT_0490, CPSIT_0594, CPSIT_0785, CPSIT_0844, CPSIT_0846, CPSIT_0019, CPSIT_0020, CPSIT_0968, CPSIT_0969, and CPSIT_0942. Ten target recombinant proteins were successfully prepared except CPSIT_0357, CPSIT_0429, CPSIT_0968 and CPSIT_0969. After immunization of BALB/c mice, the specific antibodies were prepared, and the titers were (1:32 000)-(1:64 000). Moreover, the IFA results showed that CPSIT_0844 and CPSIT_0846 were located in the inclusion membrane, while CPSIT_0461, CPSIT_0463, CPSIT_0490, CPSIT_0594, CPSIT_0785, CPSIT_0019, CPSIT_0020 and CPSIT_0942 were located in whole the inclusion bodies. In conclusion, two effector proteins localized in the Chlamydia inclusion membrane and eight effector proteins localized in the chlamydia inclusion were identified.

Key words: Chlamydia psittaci, type Ⅲ secretion system, effector proteins, inclusion membrance protein

CLC Number:

WU Haiying, TANG Ting, ZHANG Xu, PAN Yiliang, GUO Zhuang, QUAN Shufen, CHEN Lili. Localization of 10 Putative Effector Proteins Secreted via Type Ⅲ Secretion System of Chlamydia psittaci[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(6): 1354-1361.

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Localization of 10 Putative Effector Proteins Secreted via Type Ⅲ Secretion System of Chlamydia psittaci

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