Objective: To identify the possible virulence factors (VFs) of P. proliferus.

Methods: By Illumina HiSeq 4000 RNA-Seq platform, transcriptomes of adult P. proliferus worms were sequenced to predict VFs via screening the homologues of traditional VFs of parasites based on the annotations in the functional databases. Homology analysis was also performed to screen homologous genes between P. proliferus and other four Paragonimus species (i.e., P. kellicotti, P. skrjabini, P. miyazakii and P. westermani) whose transcriptomes were downloaded from the National Center for Biotechnology Information (NCBI) database, and then the differential-expressed homologous genes (DEHGs) were screened via comparisons of P. proliferus and P. kellicotti, P. skrjabini, P. miyazakii and P. westermani, respectively. Finally, an overlap of the predicted VFs and DEHGs was performed to identify possible key VFs that do not only belong to the predicted VFs but also DEHGs.

Results: A total of 1,509 genes of P. proliferus homologous to traditional VFs, including surface antigens (SAGs), secreted proteins (SPs), ATP-Binding Cassette (ABC) Transporters, actin-related proteins (ARPs), aminopeptidases (APases), glycoproteins (GPs), cysteine proteases (CPs), and heat shock proteins (HSPs), were identified. Meanwhile, homology analysis identified 6279 DEHGs among the five species, of which there were 48 DEHGs being mutually differentialexpressed among the four pairs of comparisons, such as MRP, Tuba 3, PI3K, WASF2, ADK, Nop56, DNAH1, PFK-2/FBPase2, Ppp1r7, SSP7. Furthermore, the overlap between the predicted VFs and DEHGs showed 97 genes of the predicted VFs that simultaneously belonged to DEHGs. Strikingly, of these 97 genes, only 26, including Chymotrypsin, Leucine APases, Cathepsin L, HSP 70, and so on, were higher expressed in P. proliferus while all the remaining were lower expressed than in the four other species.

Conclusion: This work provides a fundamental context for further studies of the pathogenicity of P. proliferus. Most of the predicted VFs which simultaneously belonged to DEHGs were lower expressed in P. proliferus.

目的：确定增生假单胞菌的可能毒力因子（VFs）。

方法：通过Illumina HiSeq 4000 RNA-Seq平台，对成年毕生蠕虫蠕虫的转录组进行测序，通过基于功能数据库中的注释筛选传统寄生虫VFs的同源物，预测VFs。还进行了同源性分析，以筛选增生假单胞菌与其他四个百灵纲物种（即P. kellicotti，P。skrjabini，P。miyazakii和P. westermani）之间的同源基因，其转录组从国家生物技术信息中心（NCBI）下载。 ）数据库，然后分别通过比较P. proliferus和P. kellicotti，P。skrjabini，P。miyazakii和P. westermani筛选差异表达的同源基因（DEHGs）。最后，

结果：总共有1,509个与传统VFs同源的增生毕赤酵母基因，包括表面抗原（SAG），分泌蛋白（SPs），ATP结合盒（ABC）转运蛋白，肌动蛋白相关蛋白（ARPs），氨肽酶（APases）鉴定了糖蛋白（GPs），半胱氨酸蛋白酶（CPs）和热休克蛋白（HSPs）。同时，通过同源性分析在5个物种中鉴定出6279个DEHG，其中在四对比较的MRP，Tuba 3，PI3K，WASF2，ADK，Nop56，DNAH1，PFK-2 / FBPase2等四对比较中有48个DEHG相互差异表达。 ，Ppp1r7，SSP7。此外，预测的VF和DEHG之间的重叠显示出预测的VF的97个基因同时属于DEHG。令人惊讶的是，在这97个基因中，只有26个（包括胰凝乳蛋白酶，亮氨酸APase，组织蛋白酶L，HSP 70等），

结论：这项工作为进一步研究增生假单胞菌的致病性提供了基础。同时属于DEHGs的大多数预测VFs在增生假单胞菌中表达较低。