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首页> 外文期刊>Journal of genetics >Rapid cloning and bioinformatic analysis of spinach Y chromosome-specific EST sequences
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Rapid cloning and bioinformatic analysis of spinach Y chromosome-specific EST sequences

机译：菠菜Y染色体特异性EST序列的快速克隆和生物信息学分析

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The genome of spinach single chromosome complement is about 1000 Mbp, which is the model material to study the molecular mechanisms of plant sex differentiation. The cytological study showed that the biggest spinach chromosome (chromosome 1) was taken as spinach sex chromosome. It had three alleles of sex-related X, X-m and Y. Many researchers have been trying to clone the sex-determining genes and investigated the molecular mechanism of spinach sex differentiation. However, there are no successful cloned reports about these genes. A new technology combining chromosome microdissection with hybridization-specific amplification (HSA) was adopted. The spinach Y chromosome degenerate oligonucleotide primed-PCR (DOP-PCR) products were hybridized with cDNA of the male spinach flowers in florescence. The female spinach genome was taken as blocker and cDNA library specifically expressed in Y chromosome was constructed. Moreover, expressed sequence tag (EST) sequences in cDNA library were cloned, sequenced and bioinformatics was analysed. There were 63 valid EST sequences obtained in this study. The fragment size was between 53 and 486 bp. BLASTn homologous alignment indicated that 12 EST sequences had homologous sequences of nucleic acids, the rest were new sequences. BLASTx homologous alignment indicated that 16 EST sequences had homologous protein-encoding nucleic acid sequence. The spinach Y chromosome-specific EST sequences laid the foundation for cloning the functional genes, specifically expressed in spinach Y chromosome. Meanwhile, the establishment of the technology system in the research provided a reference for rapid cloning of other biological sex chromosome-specific EST sequences.
机译：菠菜单染色体补体的基因组约为1000 Mbp，是研究植物性别分化的分子机制的模型材料。细胞学研究表明，最大的菠菜染色体（染色体1）被作为菠菜性染色体。它具有与性别相关的X，X-m和Y的三个等位基因。许多研究人员一直在尝试克隆性别决定基因，并研究菠菜性别分化的分子机制。但是，没有成功克隆有关这些基因的报道。采用了一种结合染色体显微切割和杂交特异性扩增（HSA）的新技术。将菠菜Y染色体简并寡核苷酸引物PCR（DOP-PCR）产物与雄性菠菜花的cDNA进行荧光杂交。以雌菠菜基因组为阻断剂，构建了在Y染色体上特异性表达的cDNA文库。此外，克隆了cDNA文库中的表达序列标签（EST）序列，进行了测序并分析了生物信息学。在这项研究中获得了63个有效的EST序列。片段大小在53和486bp之间。 BLASTn同源比对表明12个EST序列具有核酸的同源序列，其余为新序列。 BLASTx同源比对表明16个EST序列具有编码蛋白质的同源核酸序列。菠菜Y染色体特异的EST序列为克隆在菠菜Y染色体中特异性表达的功能基因奠定了基础。同时，该研究技术体系的建立为其他生物性染色体特异性EST序列的快速克隆提供了参考。

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