用表达序列标记法分析孤星蜱,Amblyomma americanum(蜱螨:伊蚊科)的表达基因组。

C A Hill, J A Gutierrez
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引用次数: 28

摘要

采用表达序列标记(EST)方法研究了美国Amblyomma americanum孤星蜱两个发育阶段的基因组。建立了美洲沙蚕幼虫和成虫的cDNA文库。共测序了1942条est(1462条成虫est和480条幼虫est),并利用生物信息学程序进行了分析。利用CAPII程序进行序列组装,发现幼虫est和成虫est的序列冗余度分别为11%和15%。在1942条ESTs序列中,1738条序列被认为是优质序列,其中771条序列(约44.4%)通过BLAST算法基于氨基酸身份推定鉴定。假定鉴定的序列根据其预测的基因功能进行分类。总共有967个序列,即55.6%的优质序列,与先前鉴定的基因产物具有有限的或没有蛋白质相似性。使用自动序列注释系统分析缺乏蛋白质同源性的序列,以预测蛋白质特征,如开放阅读框、信号肽、蛋白质基序和跨膜区域。在本文中,我们描述了迄今为止从一种蛛形纲动物中获得的最多数量的ESTs序列,以及随后对这些序列的详细分析。
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Analysis of the expressed genome of the lone star tick, Amblyomma americanum (Acari: Ixodidae) using an expressed sequence tag approach.

An expressed sequence tag (EST) approach was used to study the genome of two developmental stages of the lone star tick, Amblyomma americanum. cDNA libraries were constructed from the larval and adult stages of A. americanum. In total, 1942 ESTs were sequenced (1462 adult ESTs and 480 larval ESTs) and analyzed using bioinformatic programs. Contig assembly using the CAPII program revealed 11% and 15% redundancy of sequences in the larval and adult ESTs, respectively. Of the 1942 ESTs, 1738 sequences were considered quality sequences and of these, 771 or approximately 44.4% of the sequences were putatively identified based on amino acid identity using the protein Basic Local Alignment Search Tool (BLAST) algorithm. Putatively identified sequences were classified according to their predicted gene function. In total, 967 sequences, or 55.6% of the quality sequences, had limited or no protein similarity to previously identified gene products. Sequences lacking protein homology were analyzed using an automated sequence annotation system for predicted protein characteristics such as open reading frames, signal peptides, protein motifs, and transmembrane regions. In this paper we describe the sequencing of the largest number of ESTs obtained from an arachnid species to date and the subsequent detailed analysis of these sequences.

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