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Molecular cloning and its expression of trachealess gene (As-trh) during development in brine shrimp, Artemia sinica

机译:中华对虾卤虫发育过程中无气管基因(As-trh)的分子克隆及其表达

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Basic helix-loop-helix-PAS (bHLH-PAS) family transcription factors are implicated in multiple developmental and physiological regulatory processes. Herein, a full-length cDNA encoding a bHLH-PAS domain transcription factor trachealess gene (designated as As-trh) was cloned and characterized from brine shrimp (Artemia sinica) for the first time. The full-length cDNA of As-trh was 2,698 bp with a 2,319 bp open reading frame encoding a deduced protein of 772 amino acid polypeptide with a calculated molecular mass of 86.02 kDa and an isoelectric point of 5.87. Sequence alignment revealed that As-trh had high homology with other species trh gene, including the D-trh gene in Drosophila melanogaster and Bm-trh in Bombyx mori. The early and persistent expression of As-trh in the naupliar stages by whole-mount embryonic in situ hybridization and immunohistochemistry suggest that As-trh functions very early in the salt gland and may be required continuously in this tissue. Later in development, expression of As-trh begins to decrease and disappear in salt gland of the older nauplius and appears in the thoracic epipods of the sub-adult Artemia. These results indicated that As-trh might play an important role in osmoregulatiory organ development from the larvae stages through adult stages.
机译:基本的螺旋环-螺旋-PAS(bHLH-PAS)家族转录因子与多种发育和生理调节过程有关。在此,首次从盐水虾(中华卤虫)中克隆并鉴定了编码bHLH-PAS结构域转录因子无气管基因的全长cDNA(称为As-trh)。 As-trh的全长cDNA为2698 bp,具有2319 bp的开放阅读框,编码推断的772个氨基酸多肽的蛋白质,计算的分子量为86.02 kDa,等电点为5.87。序列比对表明,As-trh与其他物种trh基因具有高度同源性,包括黑腹果蝇中的D-trh基因和家蚕中的Bm-trh。通过全装载胚胎原位杂交和免疫组织化学,在无节幼体阶段As-trh的早期和持续表达表明,As-trh在盐腺中非常早地起作用,并且可能在该组织中持续需要。在发育的后期,As-trh的表达开始减少并在较老的无节幼体的盐腺中消失,并出现在亚成年卤虫的胸廓中。这些结果表明,As-trh可能在从幼虫阶段到成年阶段的渗透压器官发育中起重要作用。

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