摘要
于2007年3月,采集大连大窑湾浮筏贝类养殖区的太平洋牡蛎,抽取牡蛎的血淋巴液进行沿岸单孢子虫的研究。采用显微镜观察、聚合酶链式反应(PCR)和原位杂交(ISH)方法对血淋巴液样品进行检测。结果在显微镜下观察到牡蛎血淋巴液中存在沿岸单孢子虫(Haplosporidium costale,SSO)原生质体的类似物;提取血淋巴液基因组DNA,应用扩增沿岸单孢子虫SSUrDNA区域的引物对进行PCR扩增,产生约150bp的基因片段,经测序并将测序结果与基因库中已知序列比对分析,确定这种类似物为沿岸单孢子虫。同时,采用沿岸单孢子虫的特异性DNA探针SSO1318,对牡蛎血淋巴液中的沿岸单孢子虫进行原位杂交,结果为阳性。
Hemolymph extracted from Crassostrea gigas in Dayaowan Bay in Mar.2007 was examined.The microscope examination,polymerase chain reaction（PCR） and in situ hybridization（ISH）were applied for the study.Haplosporidium costale-like plasmodia stages were found in hemolymph by the microscope examination.DNA was extracted from the infected Hemolymph for H.costale PCR amplification.The primers were applied to amplify short regions of small subunit ribosomal DNA（SSU rDNA） from H.costale.PCR amplification of genomic DNA yielded the expected-sized product of 150 bp.This PCR product was sequenced and identified to the corresponding SSU rDNA region of H.costale.The H.costale-specific DNA probe SSO1318 was hybridized with the C.gigas parasite.
出处
《海洋环境科学》
CAS
CSCD
北大核心
2010年第3期342-345,共4页
Marine Environmental Science
基金
国家自然科学基金资助项(30470275
30070124)
国家908专项基金(9082012ZH3)
