利用RACE技术克隆并获得了银杏牻牛儿基牻牛儿基焦磷酸合成酶基因(GbGGPS基因)。银杏GbGGPS基因的cDNA全长为1 641 bp,含有1个1 176 bp的可读框,编码391个氨基酸序列。生物信息学预测GbGGPS蛋白的分子质量为42.51 ku,理论等电点为5.98,N端有叶绿体信号肽,其二级结构主要由α-螺旋和无规则卷曲组成。蛋白质同源分析表明,GbGGPS含有多聚异戊二烯基合成酶家族中保守的5个特征性结构域和2个富含天冬氨酸的区域。同源建模分析显示GbGGPS序列与薄荷GGPS蛋白的三维结构及活性位点高度相似。系统进化分析表明,银杏GGPS蛋白归属植物进化支,且与加拿大红豆杉、挪威云杉、北美冷杉的GGPS蛋白归为同一分支。
Abstract
In this study, we cloned and characterized a geranylgeranyl pyrophosphate synthase(GGPS)gene from Ginkgo biloba Linn. The full-length cDNA sequence of GbGGPS was 1 641 bp containing an open reading frame(ORF)of 1 176 bp, which encoded a 391 amino acids with a predicted molecular mass of 42.51 ku and the theoretical isoelectric point(PI)of 5.98. GbGGPS was an intro-free gene, and its deduced polypeptide contained a chloroplast-targeting signal peptide of 79 amino acids in the N terminal. The secondary structure of GbGGPS was mainly composed of alpha helix and random coil. Comparative analysis showed that GbGGPS had a high similarity to other plant GGPS proteins, and contained all the five conserved domains and functional aspartate-rich motifs of the polyprenyl synthetase family. The homology-based structural modeling showed that GbGGPS has the typical structure of Menthax piperita GGPS. Phylogenetic tree revealed that GbGGPS, TcGGDS, PaGGDS5 and AgGGDS were assigned to the same clade.
关键词
银杏萜内酯 /
牻牛儿基牻牛儿基焦磷酸合成酶(GGPS) /
蛋白质结构 /
生物信息学分析
Key words
ginkgolides /
geranylgeranyl pyrophosphate synthase(GGPS) /
protein structure /
bioinformatics analysis
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基金
收稿日期:2013-01-07 修回日期:2013-03-18
基金项目:国家林业公益性行业科研专项项目(201204403-3); 国家自然科学基金项目(31170627); 江苏省自然科学基金项目(BK2010019)
第一作者:张洪娟,博士生。*通信作者:曹福亮,教授。E-mail: zhanghj@njfu.edu.cn。
引文格式:张洪娟,谭碧玥,曹福亮. 银杏GbGGPS基因的克隆及序列分析[J]. 南京林业大学学报:自然科学版,2013,37(4):8-12.
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