Identifying genes related to choriogenesis in insect panoistic ovaries by Suppression Subtractive Hybridization.
Identifying genes related to choriogenesis in insect panoistic ovaries by Suppression Subtractive Hybridization.
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DOI：
10.1186/1471-2164-10-206
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发表时间：
2009-04-30
期刊：
影响因子：
4.4
通讯作者：
Piulachs MD
中科院分区：
文献类型：
--
作者：
Irles P;Bellés X;Piulachs MD
昆虫的卵巢管分为两类：panoistic和meroistic，后者显然是从祖先的panoistic类型进化而来的。卵子发生的分子数据实际上仅限于部分发育的卵巢。如果我们的目标是研究从panoistic到meroistic的进化过渡，应该收集panoistic子房的数据。为此，我们计划构建一个抑制性消减杂交（SSH）文库，以确定参与panoistic绒毛膜发生的基因，使用蟑螂德国小蠊为模型。本研究利用抑制消减杂交技术构建了B卵黄发生后卵巢文库，以分离与绒膜发生相关的基因。德国的。测试者文库用6至7日龄雌性的卵巢池制备，而驱动者文库用3至4日龄雌性的卵巢池制备。从SSH文库中获得了258个高质量的序列，这些序列聚成34个独特的序列，分为19个重叠群和15个单联体。使用BLAST将序列与非冗余NCBI数据库进行比较。我们发现，44%的独特序列与其他生物的已知基因具有同源序列，而56%与任何数据库条目都没有显着相似性。进行基因本体论分析，将34个序列分类为不同的功能类别。这些基因序列中的七个，代表不同的类别和过程，被选择来进行表达研究，在第一个促性腺激素周期的实时PCR。结果表明，它们主要在卵黄发生后期表达，验证了SSH技术的有效性。在其中两个对应的新基因，我们证明，他们是专门表达在细胞质中的卵泡细胞在基础卵母细胞在绒毛膜发生的时间。SSH方法已被证明在鉴定B中卵黄发生后表达的卵巢基因方面是有用的。德国的。对于大多数基因来说，都假设了与绒毛膜发生相关的功能。获得的新基因的相对高的百分比和实际上没有典型的分生卵巢的绒毛膜基因表明，在panoistic卵巢的绒毛膜形成的调节机制显着不同的分生的。
Insect ovarioles are classified into two categories: panoistic and meroistic, the later having apparently evolved from an ancestral panoistic type. Molecular data on oogenesis is practically restricted to meroistic ovaries. If we aim at studying the evolutionary transition from panoistic to meroistic, data on panoistic ovaries should be gathered. To this end, we planned the construction of a Suppression Subtractive Hybridization (SSH) library to identify genes involved in panoistic choriogenesis, using the cockroach Blattella germanica as model. We constructed a post-vitellogenic ovary library by SSH to isolate genes involved in choriogenesis in B. germanica. The tester library was prepared with an ovary pool from 6- to 7-day-old females, whereas the driver library was prepared with an ovary pool from 3- to 4-day-old females. From the SSH library, we obtained 258 high quality sequences which clustered into 34 unique sequences grouped in 19 contigs and 15 singlets. The sequences were compared against non-redundant NCBI databases using BLAST. We found that 44% of the unique sequences had homologous sequences in known genes of other organisms, whereas 56% had no significant similarity to any of the databases entries. A Gene Ontology analysis was carried out, classifying the 34 sequences into different functional categories. Seven of these gene sequences, representative of different categories and processes, were chosen to perform expression studies during the first gonadotrophic cycle by real-time PCR. Results showed that they were mainly expressed during post-vitellogenesis, which validates the SSH technique. In two of them corresponding to novel genes, we demonstrated that they are specifically expressed in the cytoplasm of follicular cells in basal oocytes at the time of choriogenesis. The SSH approach has proven to be useful in identifying ovarian genes expressed after vitellogenesis in B. germanica. For most of the genes, functions related to choriogenesis are postulated. The relatively high percentage of novel genes obtained and the practical absence of chorion genes typical of meroistic ovaries suggest that mechanisms regulating chorion formation in panoistic ovaries are significantly different from those of meroistic ones.
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DOI：
10.1006/bbrc.2001.4391
发表时间：
2001-03-02
影响因子：
3.1
作者：
Drapeau, MD
通讯作者：
Drapeau, MD
DOI：
10.1016/j.bbapap.2005.05.002
发表时间：
2005-08-01
影响因子：
3.2
作者：
Ohnishi, J;Ohnishi, E;Takahashi, T
通讯作者：
Takahashi, T
德国蟑螂亲脂蛋白受体以及脂蛋白受体的进化关系的结构和RNAi表征。
影响因子：
--
作者：
Ciudad, Laura;Belles, Xavier;Piulachs, Maria-Dolors
通讯作者：
Piulachs, Maria-Dolors
影响因子：
2.2
作者：
NORDIN, JH;BEAUDOIN, EL;LIU, XD
通讯作者：
LIU, XD
影响因子：
10.7
作者：
Albà, MM;Castresana, J
通讯作者：
Castresana, J