甘蓝型油菜PPR家族生物信息学分析与新疆野生油菜候选育性基因克隆

Bioinformatics Analysis of PPR Family in Brassica napus L. and Cloning of Candidate Restorer Gene of Xinjiang Wild Rapeseed

通过隐马尔可夫模型从甘蓝型油菜基因组中获取1079条PPR家族蛋白序列,使用拟南芥PPR家族特征模型对其进行分类,同时进行聚类、染色体分布、亚细胞定位预测、功能注释等分析。结合不育系与恢复系分子标记筛选了定位于C09染色体的PPR基因GSBRNA2T00094406001(命名为Bn PPR_C09)为新疆野生油菜潜在育性调节基因。通过分子克隆的方法从新疆野生油菜不育系1193A和恢复系1193R中分别克隆获得了长度为2514bp的c DNA序列,序列分析显示来源于1193A的Bn PPR_C09基因(Bn PPR_C09b)相对源于1193R的Bn PPR_C09a在+1725 bp位置存在单碱基缺失,造成移码突变,二者预测蛋白的生物信息学分析显示Bn PPR_C09b蛋白的N端因为移码突变导致翻译过程在+1800位置终止,后续大量功能元件缺失,该位点的突变可能决定新疆野生油菜育性。

In this study, 1079 protein sequences of PPR family were obtained from the genome of Brassica napus by Hidden Markov model. The motifs defined in the PPR family of Arabidopsis thaliana was used to classify them. The clustering, chro-mosome distribution, subcellular localization prediction, function Comments and so on were analyzed. Combining the molecular markers of the CMS line and the restorer line,the PPR gene GSBRNA2T00094406001 （named BnPPR_C09） located on C09 chromosome was screened for the potential fertility regulation gene of Xinjiang wild rapeseed. A cDNA sequence with length of 2514 bp was cloned from the CMS line 1193A of Xinjiang wild rapeseed and the restorer line 1193R by molecular cloning. Sequence analysis showed that the BnPPR_C09 gene （ BnPPR_C09b） derived from 1193A had a single base deletion at ＋ 1725 bp compared with the BnPPR_C09a derived from 1193R,resulting in frameshift mutations. The bioinformatics analysis about predicted protein showed that the frameshift mutations caused termination of the translation process at ＋ 1800 in the N - terminal of the BnPPR_C09b protein,and the subsequent deletion of a large number of functional components, the mutation may determine the fertility of Xinjiang wild rapeseed.