家蚕BmAKR基因家族的鉴定及在蚕卵中的表达分析

Genome-wide identification of the BmAKR gene family in the silkworm (Bombyx mori) and their expression analysis in diapause eggs and nondiapause eggs

摘要:

醛酮还原酶超家族(aldo-keto reductase super family, AKRs)具有广泛的底物特异性，目前尚未见对昆虫AKR基因家族成员的系统鉴定报道。本研究采用生物信息学手段，预测了家蚕(Bombyx mori) AKR基因的系统进化、理化性质、染色体定位、保守基序和基因结构，通过转录组数据和实时荧光定量聚合酶链式反应(quantitative real time polymerase chain reaction, qRT-PCR)分析了不同组织时期及不同发育状态蚕卵中BmAKR基因的表达水平，并采用Western blotting检测了蚕卵中该蛋白的表达量。本研究共鉴定出11个BmAKR基因，分布在4条不同的染色体上，都具有AKR家族特有的(α/β) 8桶保守结构，理化特征较为相似；系统进化分析显示，其可分为2个家族(AKR1和AKR2)。转录组数据分析显示，家族成员基因在不同组织时期中的表达差异较大。进一步分析发现，一部分BmAKR基因在非滞育卵的表达量显著高于滞育卵；但BmAKR1-1在滞育卵中的表达量却显著高于非滞育卵。蛋白水平的检测发现BmAKR1-1在滞育卵和非滞育卵的差异变化趋势与qRT-PCR结果一致。综上所述，本研究通过对家蚕BmAKR家族的鉴定和分析，筛选出BmAKR1-1等作为调控蚕卵发育的候选基因，以便后期对其进行深入研究。

关键词:家蚕;BmAKR基因家族;滞育卵;表达分析;系统进化

Abstract:

The aldo-keto reductase super family (AKRs) has a wide range of substrate specificity. However, the systematic identification of insect AKR gene family members has not been reported. In this study, bioinformatics methods were used to predict the phylogenetic evolution, physical and chemical properties, chromosome location, conserved motifs, and gene structure of AKR family members in Bombyx mori (BmAKR). Transcriptome data or quantitative real time polymerase chain reaction (qRT-PCR) were used to analyze the expression level of BmAKR genes during different organizational periods and silkworm eggs in different developmental states. Moreover, Western blotting was used to detect the expression level of the BmAKR in silkworm eggs. The results showed that 11 BmAKR genes were identified. These genes were distributed on 4 chromosomes of the silkworm genome, all of which had the (α/β) 8-barrel motif, and their physical and chemical characteristics were relatively similar. Phylogenetic analysis showed that the BmAKR genes could be divided into 2 subgroups (AKR1 and AKR2). Transcriptome data analysis showed that the expression of BmAKR genes were quite different in different tissues and periods. Moreover, the expression analysis of BmAKR genes in silkworm eggs showed that some genes were expressed significantly higher in nondiapause eggs than in diapause eggs; but another gene, BmAKR1-1, was expressed significantly higher in diapause eggs than in nondiapause eggs. The detection of protein level found that the difference trend of BmAKR1-1 in diapause eggs and non-diapause eggs was consistent with the results of qRT-PCR. In conclusion, BmAKR1-1 was screened out as candidates through the identification and analysis of the BmAKR genes in silkworm, which may regulate silkworm egg development is worthy of further investigation.

Key words:Bombyx mori;BmAKR gene family;diapause eggs;expression analysis;phylogenetic evolution