Transcript profiling of the meiotic drive phenotype in testis of Aedes aegypti using suppressive subtractive hybridization.
Transcript profiling of the meiotic drive phenotype in testis of Aedes aegypti using suppressive subtractive hybridization.
复制标题
使用抑制消减杂交对埃及伊蚊睾丸减数分裂驱动表型进行转录谱分析。
DOI：
10.1016/j.jinsphys.2011.05.014
复制
发表时间：
2011
影响因子：
2.2
通讯作者：
Severson,DavidW
中科院分区：
文献类型：
--
作者：
Shin,Dongyoung;Jin,Lizhong;Lobo,NeilF;Severson,DavidW
埃及伊蚊的减数分裂驱动基因与1号染色体上的性别决定位点紧密连锁，导致高度男性偏向的性别比。我们从Ae的睾丸制备cDNA文库。埃及T37菌株（驱动）和RED菌株（非驱动），并使用抑制性消减杂交技术富集T37测试特异性转录物。表达的序列标签（EST）从来自消减的T37（subT 37）文库以及每个菌株的初级文库（pT 37和pRED）的总共2784个随机选择的克隆获得。序列分析结果表明，在subT 37文库中共鉴定出171个独特基因，在3个文库中共鉴定出299个独特基因。在subT 37文库中富集的基因大多与信号转导、发育、生殖、代谢过程和细胞周期功能相关。此外，如在果蝇和小鼠中观察到的减数分裂驱动系统，这些基因中的许多与涉及调节性小GTP酶的Ras超家族的信号级联相关。这些基因中的几个的差异表达在Ae中得到验证。埃及伊蚊蛹睾丸的qRT-PCR分析。这项研究增加了我们的理解睾丸基因表达丰富的成年男性从减数分裂驱动株，以及洞察到基本的睾丸转录组在Ae。埃及人。
The meiotic drive gene in Aedes aegypti is tightly linked with the sex determination locus on chromosome 1, and causes highly male-biased sex ratios. We prepared cDNA libraries from testes from the Ae. aegypti T37 strain (driving) and RED strain (non-driving), and used suppressive subtraction hybridization techniques to enrich for T37 testes-specific transcripts. Expressed sequence tags (ESTs) were obtained from a total of 2784 randomly selected clones from the subtracted T37 (subT37) library as well as the primary libraries for each strain (pT37 and pRED). Sequence analysis identified a total of 171 unique genes in the subT37 library and 299 unique genes among the three libraries. The majority of genes enriched in the subT37 library were associated with signal transduction, development, reproduction, metabolic process and cell cycle functions. Further, as observed with meiotic drive systems in Drosophila and mouse, a number of these genes were associated with signaling cascades that involve the Ras superfamily of regulatory small GTPases. Differential expression of several of these genes was verified in Ae. aegypti pupal testes using qRT-PCR. This study increases our understanding of testes gene expression enriched in adult males from the meiotic drive strain as well as insights into the basic testes transcriptome in Ae. aegypti.