Sequence Analysis of ITS-1 and Partial HSP70 Gene of Three Cryptocaryon irritans Strains
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摘要: 经形态学观察, 初步判断从福建长乐、福鼎和宁德三都澳三地患“白点病”的牙鲆、青石斑鱼、大黄鱼鱼鳃和体表上分离收集的寄生虫为刺激隐核虫CryptocaryonirritansBrown。提取基因组DNA, 对ITS-1序列进行PCR扩增、克隆、测序和序列分析, 结果显示:3株刺激隐核虫的ITS-1序列完全一致, 与广东番禺PYH4.12株 (DQ270010) 、日本Wakayama分离株 (AB608054) 、台湾嘉义Chiayi株 (AF490381) 等序列同源性为100%, 与NCBI数据库中其他刺激隐核虫分离株的ITS-1序列的同源性为90.62%99.22%, 可以鉴定3株虫均为刺激隐核虫。进一步对刺隐核虫HSP70基因部分序列进行PCR扩增和序列分析, 结果显示:3株虫间序列差异很小, 说明福建省不同养殖模式、不同宿主得到的刺激隐核虫高度同源。Abstract: The ciliates witch isolated from Paralichthys olivaceus in Changle, Epinephelus awoara in Fuding and Pseudosciaena crocea in Sandu′ao Ningde were identified as Cryptocaryon irritans based on morphological observation.The genomic DNA of the ciliates witch was extracted, and the gene sequence of the internal transcribed spacer 1 (ITS-1) were amplified by PCR, then cloned, sequenced and analyzed.The results revealed that the ITS1nucleotide sequence of the three strains were exactly the same.Homology comparation indicated that they shared 100%identity with the gene sequence of PYH4.12strain (DQ270010) , Wakayama strain (AB608054) , Chiayi strain (AF490381) , and shared 90.62%-99.22% identity with other sequences in NCBI database, which confirmed that the three identified strain was Cryptocaryon irritans.Furthermore, partial HSP70gene sequence was amplified from the genomic DNA of the identified Cryptocaryon irritans by PCR, and only three base pairs difference were observed in the sequence among the three strains.This indicated that the Cryptocaryon irritans Strains gotten from different hosts, different farming models in Fujian were highly homologous.
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Key words:
- Cryptocaryon irritans /
- ITS /
- HSP70 /
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