Cloning and expression analysis of the DEAD-box/RNA helicase Oslaf-1 in Ovomermis sinensis
Cloning and expression analysis of the DEAD-box/RNA helicase Oslaf-1 in Ovomermis sinensis
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中华卵蝇DEAD-box/RNA解旋酶Oslaf-1的克隆及表达分析
DOI：
10.1371/journal.pone.0192101
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发表时间：
2018-02
期刊：
影响因子：
3.7
通讯作者：
Guoxiu Wang
中科院分区：
文献类型：
--
作者：
Siying Tao;Zhenlong Jiao;Guoxiu Wang
中华卵线虫是一种具有潜在防治害虫价值的线虫。线虫的寄生阶段吸收宿主血淋巴中的营养以维持其生长发育，然后在宿主出现时杀死宿主。目前，对其生殖发育，特别是其分子机制知之甚少。对生殖发育基因进行更详细的研究不仅有助于我们了解线虫性别分化的机制，而且对于成功在体外培养线虫并将其用于生物防治也很有价值。本研究采用同源克隆的方法，克隆了中华O. sinensis DEAD-box家族基因Oslaf1的全长cDNA。然后利用qRT-PCR技术检测Oslaf-1基因在不同发育阶段和组织中的表达模式，发现该基因在中华圆线虫寄生后阶段（P<0.01）和卵巢（P<0.05）中高表达。 Western blot分析显示相同的结果，即该基因与性腺发育和功能相关，但不是性腺特异性的。原位杂交进一步证明该基因在早期胚胎中广泛表达，并且主要分布在性腺区域。然而，信号主要集中在寄生前幼体的生殖原基中。 RNA干扰（RNAi）研究表明，浸泡在Oslaf-1 dsRNA中的中华稻的性别比与gfp dsRNA处理组没有统计学差异。我们的结果表明 Oslaf-1 可能在线虫的生殖系统中发挥至关重要的作用。此外，我们推测Oslaf-1基因在胚胎发育过程中发挥着重要作用，并且它在中华卵寄生前幼体的性腺中发生和发育。
Ovomermis sinensis is a potentially-valuable nematode for controlling insect pests. The parasitic stage of the nematode absorbs nutrients in its host’s hemolymph to maintain its growth development and then kills the host when it emerges. At present, little known about its reproductive development, particularly the responsible molecular mechanism. More detailed research on the genes of reproductive development will not only help us understand the mechanisms underlying sex differentiation in the nematode, but would also be valuable for successfully cultivating them in vitro and using them for biocontrol. In this study, we used the homology cloning method to clone the full-length cDNA of a DEAD-box family gene (Oslaf1) from O. sinensis. Then, using qRT-PCR technology to detect the expression pattern of the Oslaf-1 gene at different development stages and tissues, the gene was found to be highly expressed in the post-parasitic stage (P<0.01) and ovarian (P<0.05) of O. sinensis. Western blot analysis showed the same result that the gene is associated with gonadal development and function, but is not gonad-specific. In situ hybridization further demonstrated that the gene is widely expressed in early embryos and is mainly distributed in the gonadal area. However, the signal was mainly concentrated in the reproductive primordia in pre-parasitic juveniles. RNA interference (RNAi) studies revealed that the sex ratio of O. sinensis soaked in dsRNA of Oslaf-1 was not statistically different than the gfp dsRNA treated groups. Our results suggest that Oslaf-1 may play a vital role in the reproductive systems of the nematode. In addition, we speculate that the Oslaf-1 gene plays an important role during embryonic development and that it occurs and develops in the gonads of pre-parasitic juveniles of O. sinensis..
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影响因子：
1.5
作者：
L. S. Salinas;E. Maldonado;M. Macías-Silva;T. Blackwell;R. Navarro
通讯作者：
L. S. Salinas;E. Maldonado;M. Macías-Silva;T. Blackwell;R. Navarro
影响因子：
14.9
作者：
Malygin AA;Parakhnevitch NM;Ivanov AV;Eperon IC;Karpova GG
通讯作者：
Karpova GG
DOI：
10.1016/s0003-9365(88)80019-8
发表时间：
1988
期刊：
Archiv für Protistenkunde
影响因子：
--
作者：
B. R. Chaudhary;H. Singh
通讯作者：
B. R. Chaudhary;H. Singh
影响因子：
1.3
作者：
J. J. Petersen-J.
通讯作者：
J. J. Petersen-J.
棉铃虫生物防治剂中华卵蝇（线虫纲：夜蛾科）性别比例的营养依赖性模式（鳞翅目：夜蛾科）
影响因子：
1.4
作者：
Jiao, Zhenlong;Feng, Xufei;Wang, Guoxiu
通讯作者：
Wang, Guoxiu