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GPAT9基因是参与植物体中油脂代谢的重要基因之一,在甘蓝型油菜中该基因还未被深入研究。为了研究甘蓝型油菜中GPAT9基因,通过查找拟南芥At GPAT9基因(At5G60620),并以At GPAT9基因编号到Brassica Database数据库检索到白菜、甘蓝GPAT9基因序列,然后对拟南芥、白菜和甘蓝GPAT9序列进行比对。根据比对结果设计出一对引物,从甘蓝型油菜湘油15中克隆得到一条长度为1 131 bp大小的GPAT9 CDs序列,将其命名为Bn GPAT9。Bn GPAT9基因共编码376个氨基酸,相对分子量为43.25 k D,等电点为8.97,不稳定系数为46.46;Bn GPAT9编码的蛋白有三个跨膜结构域,没有信号肽;该蛋白的二级结构中α螺旋为38.03%,β折叠为21.28%,β转角为9.04%,无规则卷曲为31.65%。分析表明:无论核苷酸序列还是氨基酸序列聚类均显示Bn GPAT9与拟南芥、白菜和甘蓝GPAT9亲缘关系近;Bn GPAT9具有溶血磷脂酰基转移酶(LPLAT)的结构域LPCAT1-like和GPAT9特有的活性位点。从序列分析、理化性质分析、二级结构预测、亚细胞定位和功能结构域分析等都表明克隆到的基因为甘蓝型油菜Bn GPAT9基因,属于GPAT基因家族成员。
The GPAT9 gene is one of the important genes involved in lipid metabolism in plants. The gene has not been further studied in Brassica napus. In order to study the GPAT9 gene in Brassica napus, the GPAT9 gene sequence of Brassica campestris and Brassica campestris was obtained by searching the At GPAT9 gene (At5G60620) and the At GPAT9 gene number to the Brassica Database. Then GPAT9 was expressed in Arabidopsis, Brassica campestris and Brassica campestris Sequence alignment. A pair of primers was designed based on the alignment results. A GPAT9 CDs sequence of 1 131 bp in length was cloned from the Brassica napus Linyou 15 and named as Bn GPAT9. The Bn GPAT9 gene encodes a protein of 376 amino acids with a relative molecular mass of 43.25 kD, an isoelectric point of 8.97 and an instability coefficient of 46.46. The protein encoded by Bn GPAT9 has three transmembrane domains and no signal peptide. The secondary structure of the protein The α-helix was 38.03%, the β-sheet was 21.28%, the β-angle was 9.04% and the irregular curl was 31.65%. The results showed that Bn GPAT9 was closely related to GPAT9 in Arabidopsis thaliana, Brassica campestris and Brassica oleracea both in terms of nucleotide sequence and amino acid sequence clustering. Bn GPAT9 had the LPCAT1-like and GPAT9-specific domains of lysophosphatidyl transferase (LPLAT) Of the active site. Sequence analysis, physical and chemical properties analysis, secondary structure prediction, subcellular localization and functional domain analysis showed that the cloned gene is Brassica napus Bn GPAT9 gene, belonging to the GPAT gene family members.