家蚕孤雌生殖差异脂蛋白30K lipoprotein precursor基因的克隆与生物信息学分析

应用二维凝胶电泳(2-DE)技术分离家蚕孤雌生殖蚁蚕总蛋白,并和正常蚁蚕总蛋白进行比较,得到46个可能与家蚕孤雌生殖相关的差异蛋白点。对这些差异蛋白点进行胶内酶解后基质辅助激光解吸电离串联飞行时间质谱(MALDI-TOF-TOFMS)分析,经数据库搜索鉴定,确定其中一个差异蛋白点为脂蛋白30K lipoprotein precursor(30K LPP)。通过5′-RACE技术克隆到30KLPP脂蛋白的全长cDNA序列。生物信息学分析显示该差异蛋白基因全长917bp,编码261个氨基酸,理论分子质量为30.013kD,等电点为7.193;结构域和三级结构预测显示该蛋白包含1个Lipoprotein-11结构域和12个β-折叠。预测其信号肽概率为1.000,定位在1~20aa区域,即可能存在信号肽。细胞定位预测显示该蛋白可能分泌到胞外(SP值为0.934)。分析结果有助于进一步研究30KLPP脂蛋白结构与功能的关系。

Cloning and Bioinformatics Analysis of a Parthenogenesis-related 30K Lipoprotein Precursor Gene in Bombyx mori

Parthenogenesis is a phenomena that female sex cell is unfertilized and developed into individual directly. Comparing with total proteins of Bombyx mori hatched larva from sexual reproduction, 46 kinds of parthenogenesis-related protein spots were obtained in total proteins of hatched larva from parthenogenetic reproduction by two-dimensional gel electrophoresis (2-DE) technology. These differential protein spots were digested in the gel and identified by MALDI-TOF-TOF MS. One of these proteins was a 30K lipoprotein precursor (30K LPP) by searching database. The full-length cDNA sequence of 30K LPP was cloned using 5 -RACE technique. Bioinformatics analysis showed that 30K LPP gene, containing 917 nucleotides, encoded 261 amino acids with a predicted molecular weight of 30.013 kD and pI of 7.193. Additionally, the sequence of this protein harbored a conserved domain, Lipoprotein_11, and 12 beta-folds. Prediction of signal peptide showed that the probability of signal peptide was 1.000 and it was located in the region 1~20aa, indicating that this protein might exist signal peptide. Prediction of subcellular location suggested that this protein could be excreted from the cells (the SP value is 0.934). The present experiment results will provide favorable reference for further research on the relationship between structure and function of 30K LPP .