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首页> 外文期刊>DNA and Cell Biology >Mouse dynein axonemal intermediate chain 2: cloning and expression.
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Mouse dynein axonemal intermediate chain 2: cloning and expression.

机译:小鼠动力蛋白轴突中间链2:克隆和表达。

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摘要

Ovarian follicular development is a complex process. Investigation of the mechanisms regulating the initiation of follicular growth, and the growth and differentiation of preantral follicles is of great interest. In an effort to clone follicular development-related genes, we selected a partial cDNA fragment by differential display reverse-transcription PCR using total RNA extracted from 5-day-old and 10-day-old mouse ovaries, and its open reading frame was obtained by rapid amplification of cDNA ends. Sequencing showed that the fragment is the mouse dynein axonemal intermediate chain 2 gene (Dnaic2), which has an 87% homology with human DNAI2, a candidate gene for primary ciliary dyskinesia. Northern and western analyses indicate that Dnaic2 produces an approximate 3 kb mRNA that is translated into an approximate 70 kDa protein. The mRNA is predominantly expressed in mouse ovary, testis, and lung. In mouse ovaries, Dnaic2 mRNA was detected at high levels in vivo on day 10, with a subsequent decrease on days 15 and 20, in adult and old ovaries. However, Dnaic2 expression was weak on day 5. Dnaic2 protein was localized on the surface of the oocyte. No obvious fluorescence signal was detected in primordial and primary follicles, while strong signals were detected on the oocyte surface of secondary and antral follicles, in particular for secondary follicles in day 10. These data suggest that Dnaic2 plays a role in ovarian follicular development.
机译:卵巢卵泡发育是一个复杂的过程。研究调节卵泡生长的起始机制,以及窦前卵泡的生长和分化的机制引起了极大的兴趣。为了克隆与卵泡发育相关的基因,我们使用从5天和10天大的小鼠卵巢中提取的总RNA,通过差异显示逆转录PCR选择了部分cDNA片段,并获得了其开放阅读框通过快速扩增cDNA末端。测序表明该片段是小鼠动力蛋白轴突中间链2基因(Dnaic2),与人DNAI2(原发性睫状运动障碍的候选基因)具有87%的同源性。北方和西方的分析表明,Dnaic2产生大约3 kb的mRNA,并翻译成大约70 kDa的蛋白质。该mRNA主要在小鼠卵巢,睾丸和肺中表达。在小鼠卵巢中,第10天在体内检测到高水平的Dnaic2 mRNA,随后在成年和老年卵巢中在第15天和第20天降低Dnaic2 mRNA。但是,Dnaic2表达在第5天很弱。Dnaic2蛋白位于卵母细胞表面。在第10天,在原始卵泡和初级卵泡中未检测到明显的荧光信号,而在次级卵泡和肛门卵泡的卵母细胞表面(尤其是次级卵泡)的卵母细胞表面检测到强信号。这些数据表明Dnaic2在卵巢卵泡发育中起作用。

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