摘要： 为了进一步研究甘蓝型油菜GPDH基因，通过从拟南芥GPDH基因与甘蓝型油菜的共线性分析发现：拟南芥GPDH基因（At2g41540）在甘蓝型油菜A基因组和C基因组中有4个拷贝，分别为：GSBRNA2T00132785001、GSBRNA2T00024258001、GSBRNA2T00058395001、GSBRNA2T00019118001。从GenBank分别下载了它们的DNA序列和cDNA序列，cDNA序列依次命名为GPDH1、GPDH2、GPDH3、GPDH4。根据序列信息设计了4对不同的引物，从甘蓝型油菜湘油15号cDNA全长序列中扩增出了4条片段，分别命名为BnGPDH1、BnGPDH2、BnGPDH3、BnGPDH4。利用生物信息学软件和在线系统进行核苷酸序列分析和蛋白进化分析。结果显示：4条片段大小依次为1 395，1 395，1 389，1 536 bp。其中，BnGPDH4翻译成氨基酸时序列中间出现终止密码子，不能合成有效蛋白。3个BnGPDH蛋白的氨基酸数目为464，464和462个，理论分子量为51.584 4~51.829 7 kDa，二级结构α螺旋和无规则卷曲所占比例在42%左右，其次是延伸链所占比例约为15%。三者的跨膜结构总体来说是相同的。3个蛋白都由保守结构GpsA、NADB_Rossmann superfamily和NAD_Gly3P_dh_C superfamily组成，属于GPDH超基因家族。同源建模分析表明3个蛋白均能适用甘油3磷酸脱氢酶（Glycerol-3-phosphate dehydrogenase，GPDH）模型。对其进行亚细胞定位预测在细胞质膜的概率为79.0%。从序列分析、核苷酸聚类、理化性质预测、二级、三级结构预测、亚细胞定位预测、氨基酸聚类分析得出结论，它们具有GPDH基因家族的特征，为甘蓝型油菜GPDH基因家族中的成员。

关键词: GPDH, 甘蓝型油菜, 生物信息学分析

Abstract: To further study on GPDH of Brassica napus,through the collinearity analysis of the GPDH from Arabidopsis thaliana and Brassica napus:there are 4 copies of GPDH of A genome and C genome in the Brassica napus called as GSBRNA2T00132785001,GSBRNA2T00024258001,GSBRNA2T00058395001,GSBRN-A2T00019118001.Four cDNA were downloaded from GenBank,are named as:GPDH1,GPDH2,GPDH3 and GPDH4. Four pairs of primers were designed according to the sequence informations,and four nucleotide fragments were amplified from Xiangyou 15 full-length cDNA,designated as BnGPDH1,BnGPDH2,BnGPDH3,BnGPDH4. The result of nucleotide sequences and protein structure analysis by bioinformatics software and online system showed:Four CDS sequences length were 1 395,1 395,1 389,1 536 bp respectively.When BnGPDH4 is translated into the amino acid sequence,we found a stop codon in the middle of BnGPDH4. Which isn't an active protein.The number of amino acid of three other BnGPDH are 464,464 and 462 respectively.Secondary structure are consistant of the alpha helix (42%),random coil (42%)and extended chain (15%).The transmembrane structure of the three proteins was uniform.Their conservative domains consist of GpsA,NADB_Rossmann superfamily and NAD_Gly3P_dh_C superfamily,which belong to the GPDH superfamily.Homology modeling analysis showed that they were of model of Glycerol-3-phosphate dehydrogenase and located in cytoplasm by subcellular localization analysis.In terms of sequence composition,phylogenic tree,physico-chemical property,secondary structure and tertiary structure and subcellular localization,they have the characteristics of GPDH superfamily.They should be the members of GPDH superfamily.

Key words: GPDH, Brassica napus L., Bioinformatics analysis

中图分类号: 

• Q78

• S635.03