Shin Jin Ho, Joo Han Soo, Lee Won Hyung, Seok Ho Bong, Calsamig Maria, Pijoan Carlos, Molitor Thomas W
Department of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul 55108, USA.
J Vet Med Sci. 2003 Apr;65(4):501-9. doi: 10.1292/jvms.65.501.
A virus-like cytopathic agent isolated from swine farms with a history of recurrent abortion episodes was investigated. We employed a differential display reverse transcription-polymerase chain reaction (ddRT-PCR) to obtain genetic information of the cytopathic agent. Partial nucleotide sequence (527 bp) obtained from differentially displayed PCR fragments showed 88.7% similarity with the 23S rRNA gene of Mycoplasma hyopneumoniae. Unexpectedly, the 5' portion (1-333 bp) of the sequence shared 96.1% similarity with 5' untranslated region (UTR) of human prostate tumor inducing gene 1 (PTI-1). Cytopathic effects and extranuclear DNA fluorescence were no longer observed when BM-cyclin was added in the culture medium, suggesting that BM-cyclin sensitive mycoplasma-like organisms caused the cell death. Further evidence supporting the cytopathic agent as a mycoplasma-like organism was obtained by the capability of (3)H-thymidine and (3)H-uridine incorporation, a single peak in buoyant density gradient profile (1.20-1.24 g/ml), and ultrastructural morphology. Unlike M. hyopneumoniae, the organism was not propagated in Friis medium. Nucleotide sequence of 16S rRNA obtained from the cytopathic agent showed 0.8-1.0% divergences with other M. hyorhinis strains, suggesting that the newly isolated cytopathogenic swine mycoplasma was a variant form of M. hyorhinis. Striking homology between a portion of the 23S rRNA gene of M. hyorhinis and 5' UTR of human PTI-1 implicated that M. hyorhinis might potentially be related to the evolution of human PTI-1.
对从有反复流产病史的猪场分离出的一种病毒样细胞病变因子进行了研究。我们采用差异显示逆转录-聚合酶链反应(ddRT-PCR)来获取该细胞病变因子的遗传信息。从差异显示PCR片段获得的部分核苷酸序列(527 bp)与猪肺炎支原体的23S rRNA基因显示出88.7%的相似性。出乎意料的是,该序列的5'部分(1-333 bp)与人前列腺肿瘤诱导基因1(PTI-1)的5'非翻译区(UTR)有96.1%的相似性。当在培养基中添加BM-细胞周期蛋白时,细胞病变效应和核外DNA荧光不再出现,这表明对BM-细胞周期蛋白敏感的支原体样生物导致了细胞死亡。通过(3)H-胸腺嘧啶核苷和(3)H-尿苷掺入能力、浮力密度梯度图谱中的单峰(1.20-1.24 g/ml)以及超微结构形态,获得了支持该细胞病变因子为支原体样生物的进一步证据。与猪肺炎支原体不同,该生物在弗里斯培养基中不能繁殖。从该细胞病变因子获得的16S rRNA核苷酸序列与其他猪鼻支原体菌株有0.8-1.0%的差异,表明新分离的细胞病变性猪支原体是猪鼻支原体的一种变异形式。猪鼻支原体23S rRNA基因的一部分与人PTI-1的5' UTR之间的显著同源性表明,猪鼻支原体可能与人类PTI-1的进化潜在相关。