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来自法国大西洋沿岸的太平洋牡蛎(Crassostrea gigas)中的单孢子虫病。

Haplosporidiosis in the Pacific oyster Crassostrea gigas from the French Atlantic coast.

作者信息

Renault T, Stokes N A, Chollet B, Cochennec N, Berthe F, Gérard A, Burreson E M

机构信息

IFREMER, Laboratoire de Génétique et Pathologie, La Tremblade, France.

出版信息

Dis Aquat Organ. 2000 Sep 28;42(3):207-14. doi: 10.3354/dao042207.

Abstract

Two cases of haplosporidian infection occurred during 1993 in Pacific oysters Crassostrea gigas from the French Atlantic coast. The localization and ultrastructure of the plasmodia are described. In situ hybridization of infected tissue sections was conducted with DNA probes for oyster-infecting haplosporidians. The Haplosporidium nelsoni-specific DNA probe MSX1347 hybridized with the C. gigas parasite, and the H. costale-specific probe SSO1318 did not hybridize. Total genomic DNA was extracted from the infected tissue sections for polymerase chain reaction (PCR) amplification of the haplosporidian. PCR amplifications with H. nelsoni-specific primers and with 'universal' actin primers did not yield the expected products of 573 and 700 bp, respectively. A series of primers was designed to amplify short regions of small subunit ribosomal DNA (SSU rDNA) from most haplosporidians. The primers encompass a highly variable region of the SSU rDNA and did not amplify oyster DNA. PCR amplification of the infected C. gigas genomic DNA with these primers yielded the expected-sized product from the primer pair targeting the shortest region (94 bp). This PCR product was sequenced and it was identical to the corresponding SSU rDNA region of H. nelsoni.

摘要

1993年,法国大西洋沿岸的太平洋牡蛎(Crassostrea gigas)中出现了两例单孢子虫感染病例。本文描述了疟原虫的定位和超微结构。使用针对感染牡蛎的单孢子虫的DNA探针,对感染组织切片进行原位杂交。尼尔森单孢子虫特异性DNA探针MSX1347与太平洋牡蛎寄生虫杂交,而科氏单孢子虫特异性探针SSO1318未杂交。从感染组织切片中提取总基因组DNA,用于单孢子虫的聚合酶链反应(PCR)扩增。使用尼尔森单孢子虫特异性引物和“通用”肌动蛋白引物进行PCR扩增,分别未产生预期的573和700 bp产物。设计了一系列引物,用于扩增大多数单孢子虫的小亚基核糖体DNA(SSU rDNA)的短区域。这些引物涵盖了SSU rDNA的一个高度可变区域,不会扩增牡蛎DNA。用这些引物对感染的太平洋牡蛎基因组DNA进行PCR扩增,从靶向最短区域(94 bp)的引物对中得到了预期大小的产物。对该PCR产物进行测序,结果与尼尔森单孢子虫的相应SSU rDNA区域相同。

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