Ko Y T, Ford S E, Fong D
Department of Biological Sciences, Rutgers University, Piscataway, New Jersey, USA.
Mol Mar Biol Biotechnol. 1995 Sep;4(3):236-40.
The small subunit ribosomal RNA (SSU rRNA) gene of the oyster parasite Haplosporidium costale was characterized from spore DNA by polymerase chain reaction (PCR) and molecular cloning. Sequence analysis showed that identical clones were obtained from separate batches of spore samples. The gene is 1791 nucleotides in size. It has 84.5% sequence similarity to that of a related oyster parasite, Haplosporidium nelsoni, 71.8% similarity to that of its oyster host, Crassostrea virginica, and 75.4% similarity to that of another oyster parasite, Perkinsus marinus. Among the variable regions of these SSU rRNA genes, H. costale-specific primers were designed and used to confirm parasite identity by the PCR technique. A common 150 base pair amplification product was obtained from DNA samples of H. costale spore DNA, DNA prepared from tissue sections of oysters infected with H. costale plasmodia, and the H. costale SSU rRNA clone. There was no detectable product from DNA samples isolated from tissue sections of oysters infected with H. nelsoni plasmodia.
通过聚合酶链反应(PCR)和分子克隆技术,从牡蛎寄生虫“科氏单孢子虫”(Haplosporidium costale)的孢子DNA中鉴定出小亚基核糖体RNA(SSU rRNA)基因。序列分析表明,从不同批次的孢子样本中获得了相同的克隆。该基因大小为1791个核苷酸。它与相关的牡蛎寄生虫“尼尔森单孢子虫”(Haplosporidium nelsoni)的序列相似度为84.5%,与其牡蛎宿主“弗吉尼亚巨蛎”(Crassostrea virginica)的相似度为71.8%,与另一种牡蛎寄生虫“玛丽娜派琴虫”(Perkinsus marinus)的相似度为75.4%。在这些SSU rRNA基因的可变区域中,设计了科氏单孢子虫特异性引物,并用于通过PCR技术确认寄生虫的身份。从科氏单孢子虫孢子DNA的DNA样本、从感染科氏单孢子虫疟原虫的牡蛎组织切片制备的DNA以及科氏单孢子虫SSU rRNA克隆中获得了一个常见的150个碱基对的扩增产物。从感染尼尔森单孢子虫疟原虫的牡蛎组织切片分离的DNA样本中未检测到产物。