黄鳝性腺差异表达基因的筛选和克隆

Screening and Cloning of Differentially Expressed Genes from Gonad of Monopterus albus

运用ACP-DDRT-PCR技术筛选和克隆了黄鳝不同发育时期性腺组织的差异表达基因。结果显示:利用2个随机ACP引物筛选到2个表达差异显著的基因,并对这2条差异表达片段进行克隆、测序分析,获得这2个基因的部分cDNA序列,长度分别为408 bp和421 bp。同源性分析结果显示其中一个基因与黄鲈卵巢cDNA文库中的一个基因(GeneBank No.GO657149.1)高度同源;而另一个基因无同源性序列,视为新报道的基因。进一步半定量RT-PCR检测,结果显示:这两个基因在黄鳝卵巢和间期性腺中的表达差异显著,因此推测其在黄鳝性腺发育以及性逆转过程中起着重要作用。

Using annealing control primer （ACP） based differential display reverse transcription polymerase chain reaction （ACP-DDRT-PCR） to screen and clone differentially expressed genes （DEGs） from gonad in ovary at stage IV and ovotestis stages of Monopterus albus. Using two arbitrary ACP primers to identify two DEGs, and then these two were cloned and sequence analyzed to obtained parts of cDNA sequences of these two genes, and the length of them were 408bp and 421 bp, respectively. The homologous analysis revealed that one of the genes has highly homologous to one of the genes in eDNA library of yellow perch ovarian （GeneBank No.GO657149.1）; while the other gene has non- homologous sequence, which is a new reported gene. Further Semi-quantitative RT-PCR test results revealed that there was a significantly difference between expressions of these two genes in ovary at stage IV and ovotestis of Monopterus a/bus. So it supposed that these two genes play important roles in the gonadal development and sex-reversal of Monopterus albus.