家蚕第2白卵近等基因系差异表达基因的筛选

Screening of Differentially Expressed Genes from the Near-isogenic Line of w-2 in the Silkworm,Bombyx mori

为了解与家蚕第2白卵(w-2)性状形成相关的差异表达基因信息,以家蚕正常型黑卵及其第2白卵近等基因系的转色期蚕卵为材料,构建抑制消减杂交(SSH)文库,筛选差异表达基因。对SSH文库中部分克隆的测序分析表明,该文库对差异表达基因的富集性较好。随机挑选SSH文库中的300个克隆制作家蚕cDNA芯片,对家蚕正常型黑卵及第2白卵近等基因系转色期蚕卵进行检测,获得11个差异表达基因。对这11个差异表达基因进行实时荧光定量RT-PCR验证分析,其结果与芯片数据分析结果趋势一致,在正常型黑卵与第2白卵近等基因系之间,这些基因的表达差异为0.1倍至数千倍。

In this study, the library of suppression subtractive hybridization （SSH） from wild type strain Jingsong A and its w-2 near-isogenic line was constructed to explore the differentially expressed genes of eggs in the silkworm, Bombyx mori. Partial clone sequencing to SSH library indicated that the obtained library had a good enrichment of the differentially expressed genes. 300 clones were selected randomly from SSH library to produce cDNA microarray, and 11 differentially expressed genes were identified from the cDNA microarray analysis to silkworm eggs at the pigmentation stage from both normal black egg strain and w-2 near-isogenic line. Results from quantitative analysis of real-time RT-PCR to the 11 differentially expressed genes were consistent with cDNA microarray data, showing differential expression levels ranging from 0.1 to thousands of times between the normal black egg strain and the w-2 near-isogenic line.