已知卵巢肿瘤基因(ovarian tumor gene,otu)在果蝇(Drosophila melanogaster)卵巢发育过程中发挥极其重要的作用,该基因突变会扰乱卵子形成的正常过程。为探究家蚕(Bombyx mori)是否具有类似果蝇生殖发育相关基因otu的同源基因,以及基因存在的可变剪接形式,在电子克隆的基础上,应用RT-PCR从家蚕精巢组织中获得了4条不同长度的Bmotu cDNA片段(GenBank登录号:HQ831341,HQ831342,HQ999998,HQ831343),其中3条由于无义突变导致翻译提前终止;而从卵巢组织中仅获得了1条Bmotu cDNA片段,该片段序列与精巢中扩增的登录号为HQ831343片段序列的5'端相同。结果表明Bmotu基因存在可变剪接,且雌、雄之间的剪接方式可能存在差异。生物信息学分析发现Bmotu基因编码蛋白与果蝇Otu的结构类似,含有半胱氨酸蛋白酶结构域(Otu结构域)、Tudor结构域、脯氨酸基序。研究结果有助于进一步探讨家蚕生殖发育机制。
Ovarian tumor gene(otu) plays a pivotal role in ovary development of Drosophila and mutations in otu gene disrupt the normal process of oogenesis.To investigate whether silkworm(Bombyx mori) has a homologous gene similar to germ line development-related gene otu in Drosophila and the isoforms produced by alternative splicing of the homologous gene,four Bmotu cDNA segments with varied lengths from silkworm testis were obtained using RT-PCR based on in silico cloning(GenBank accession No.HQ831341,HQ831342,HQ999998 and HQ831343).Three of them bear stop codons which would terminate translation prematurely due to nonsense mutation.Nevertheless,only one Bmotu cDNA fragment was obtained from ovary.Sequence of this fragment is identical to the 5′ end of fragment HQ831343 which was amplified from testis.These results indicated that Bmotu gene has alternative splicing and that diverse alternatively spliced isoforms exist in male and female silkworms.Bioinformatics analysis discovered that proteins encoded by Bmotu are similar to Drosophila Otu in structure which contains a cysteine proteinase domain(Otu domain),a Tudor domain and a proline-rich motif.The obtained data are conducive to further investigation on mechanism of silkworm germ line development.