Studies on Murine Retrovirus-Related Sequences in the Schistosome. DNA

血吸虫中鼠逆转录病毒相关序列的研究。

• 批准号: 63480149
• 负责人: IRIE Yuji
• 金额: $ 4.35万
• 依托单位: University of Tsukuba
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1990
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63480149/
• 关键词: Schistosomes; DNA; Retrovirus-related sequence; Southern blot; Western blot; ABC method; PCR method

Southern hybriaizdtion of the DNA extracted from adults of Schistosoma japonicum and S. mansoni demonstrated the presence of sequences homologous to the env gene of mouse ecotropic and xenotropic retroviruses. DNA sequences homologous to the gag and pol regions of the ecotropic murine leukemia virus, and to the mouse intracisternal A particle and endogenous type C retrovirus were also detected in the DNA of S. japonicum adults and S. mansoni eggs.Antibodies against the retrovirus envelope glycoprotein (gp70) of mouse xenotropic retrovirus, BALB virus 2 (Bv2) reacted with the adult worms of S. japonicum and S. mansoni. This reaction was completely inhibited after adsorption of the antibodies with virions of retrovirus. The reactive schistosome antigen was located in the subsegumental layer of adult male fluke and in the vitelline gland of the adult female of S. japonicum and S. Bmasoni. Proteins extracted from both parasites were examined by immunoblot analysis. Anti-Bv2 gp70 antiserem … More reacted with those proteins from both schistosomes and the band patterns were different among sexes and species.Hybridization with a clone SmE15 DNA from S. Bmasoni miracidia as a probe showed no signal in the DNAs from S. Bmansoni adults, indicating these sequences deleted in adults. Hybridization experiments using the probes SmF25 and SmM51 which are 1.3 and 2.2kb fragments cloned from s. mansoni adult females and males respectively, demonstrated no signal to DNA from S. mansoni miracidia. These data suggested the existence of stagespecific DNA sequences in S. mansoni.Repetitive sequences in the host genome such as mouse type l Alu sequence (Bl), mouse type 2 Alu sequence (B2) and mo-2 sequence, a mouse minisatellite, were detected in the DNAs from adults and eggs of S. japonicum and dggs of S. mansoni. Almost all of the sequences described above were absent in the DNAs of S. mansoni adults. The DNA fingerprints of schistosomes, using the mo-2 sequence, were indistinguishable from each other and resembled those of their murine hosts. Morever, the mo-2 sequence was hypermethylated in the DNAs of schistosomes and its amount was variable in them. These facts indicate that host-related sequence are actually present in schistosomes and that the mo-2 repetitive sequence exists probably in extra-chromosome.Polymerase chain reaction using the primer sets of gag region of retrovirus revealed the amplification of gag -related sequences in the DNAs of S. japonicum male adults and S. mansoni eggs. Less

用日本血吸虫成虫和日本血吸虫成虫DNA进行Southern杂交。Mansoni证明存在与小鼠嗜亲性和异嗜性逆转录病毒的env基因同源的序列。在S. japonicum成虫和S.抗小鼠异嗜性逆转录病毒BALB病毒2型（Bv 2）包膜糖蛋白（gp 70）的抗体与曼氏血吸虫蠕虫发生反应。japonicum和S. mansoni用逆转录病毒的病毒粒子吸附抗体后，该反应被完全抑制。反应性溶酶体抗原定位于雄体亚节层和雌体卵黄腺中。japonicum和S.布马索尼从两种寄生虫中提取的蛋白质通过免疫印迹分析进行检查。抗Bv 2 gp 70抗体 ...更多信息 结果表明，SmE 15与两种染色体上的蛋白质均发生反应，且不同种、不同性别的SmE 15 DNA有不同的带型。Bmasoni毛蚴作为探针在S. Bmansoni成虫，表明这些序列在成虫中缺失。用从日本血吸虫中克隆的1.3kb和2.2kb的SmF 25和SmM 51探针进行杂交实验。mansoni成虫雌虫和雄虫对S.曼氏毛蚴。这些数据表明，在S.在成虫和卵的DNA中检测到小鼠1型Alu序列（B1）、小鼠2型Alu序列（B2）和小鼠小卫星mo-2序列等宿主基因组中的重复序列。japonicum和S. mansoni上述序列几乎都不存在于S.曼索尼成人。使用mo-2序列，染色体的DNA指纹彼此难以区分，并且与其小鼠宿主的DNA指纹相似。此外，在线粒体DNA中mo-2序列高甲基化，其数量是可变的。用逆转录病毒gag区的引物对进行聚合酶链反应，结果显示在S. japonicum雄成虫和S.曼索尼鸡蛋。少

项目成果

Iwamura Y, Irie Y, Kominami R, Nara T, and Yasuraoka K.: "Existence of host-related DNA sequences in schistosome genome" Paraditology. 102.

Iwamura Y、Irie Y、Kominami R、Nara T 和 Yasuraoka K.：“血吸虫基因组中宿主相关 DNA 序列的存在”Paraditology。

Takeshi Nara: "Dynamic changes of DNA sequences in <Schistosoma>___ー <mansoni>___ー in the course of development" Parasitology. 100. 241-245 (1990)

奈良武 (Takeshi Nara)：“<血吸虫>____ <曼索尼>____ 发育过程中 DNA 序列的动态变化”《寄生虫学》100. 241-245 (1990)。

Manami,Tanaka: Parasitology.

田中真奈：寄生虫学。

Nara T, Iwamura Y, Tanaka M, Irie Y, and Yasuraoka K.: "Dynamic changes of DNA sequences in the course of development" Parasitology. 100. 241-245 (1990)

Nara T、Iwamura Y、Tanaka M、Irie Y 和 Yasuraoka K.：“发育过程中 DNA 序列的动态变化”寄生虫学。

Manami Tanaka: "Integration and expression of murine retrovirus-related sequences in schistosomes" Parasitology. 99. 31-38 (1989)

Manami Tanaka：“血吸虫中鼠逆转录病毒相关序列的整合和表达”寄生虫学。