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曼氏血吸虫:运用消减克隆策略在抗寄生虫的光滑双脐螺中寻找限制性片段长度多态性

Schistosoma mansoni: use of a subtractive cloning strategy to search for RFLPs in parasite-resistant Biomphalaria glabrata.

作者信息

Miller A N, Ofori K, Lewis F, Knight M

机构信息

Biomedical Research Institute, Rockville, Maryland 20852, USA.

出版信息

Exp Parasitol. 1996 Dec;84(3):420-8. doi: 10.1006/expr.1996.0130.

Abstract

A subtractive cloning strategy has been applied for the identification of two cDNA clones whose corresponding transcripts were elevated in Schistosoma mansoni-resistant (BS-90) compared to susceptible (M-line) snails. Clone pBS11 encoded a 1.9-kb transcript that was more elevated compared to a 500-bp transcript encoded by clone pBS12. Consequently, more attention was focused on the molecular characterization of clone pBS11. Results showed that the transcript encoded by this clone was expressed in the albumen gland and was developmentally regulated. Sequence analysis of pBS11 demonstrated the presence of an open reading frame that corresponded to a novel Biomphalaria glabrata albumen gland gene product. Comparative Southern analysis of the resistant and susceptible snail lines using pBS11 as probe indicated the presence of a BamHI and EcoRI RFLP between the two strains.

摘要

一种消减克隆策略已被用于鉴定两个cDNA克隆,与易感(M系)蜗牛相比,其相应转录本在曼氏血吸虫抗性(BS-90)蜗牛中表达上调。克隆pBS11编码一个1.9 kb的转录本,与克隆pBS12编码的500 bp转录本相比,其表达上调更为明显。因此,更多的注意力集中在克隆pBS11的分子特征上。结果表明,该克隆编码的转录本在蛋白腺中表达,并受发育调控。pBS11的序列分析表明存在一个开放阅读框,对应于一种新的光滑双脐螺蛋白腺基因产物。用pBS11作为探针,对抗性和易感蜗牛品系进行比较Southern分析,结果表明两个品系之间存在BamHI和EcoRI限制性片段长度多态性(RFLP)。

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