象草PAL基因克隆及其蛋白质结构与功能预测

doi:10.11733/j.issn.1007-0435.2016.01.019

草地学报 ›› 2016, Vol. 24 ›› Issue (1): 137-145.DOI: 10.11733/j.issn.1007-0435.2016.01.019

象草PAL基因克隆及其蛋白质结构与功能预测

陈卫卫¹, 唐然¹, 吴昱煜¹, 李菲¹, 张向前¹, 解新明^1,2

1. 华南农业大学林学与风景园林学院, 广东广州 510642;
2. 广东省草业工程技术研究中心, 广东广州510642

收稿日期:2014-11-28 修回日期:2015-04-03 出版日期:2016-02-15 发布日期:2016-04-26
通讯作者: 解新明
作者简介:陈卫卫(1990-),女,硕士研究生,主要从事牧草基因功能研究,E-mail:1039553248@qq.com;同等贡献作者,唐然(1988-),男,博士研究生,主要从事牧草基因功能研究,E-mail:tr880213@163.com
基金资助:
国家自然科学基金(31272491,30972138);教育部高等学校博士学科点专项科研基金(20124404110011)资助

Cloning of a PAL Gene of Pennisetum purpureum and Prediction of Its Protein Structure and Function

CHEN Wei-wei¹, TANG Ran¹, WU Yu-yu¹, LI Fei¹, ZHANG Xiang-qian¹, XIE Xin-min^1,2

1. Department of Grassland Science, College of Forestry and Landscape Archittecture, South China Agricultural University, Guangzhou, Guangdong Province 510642, China;
2. Guangdong Engineering Research Center for Grassland Science, Guangzhou, Guangdong Province 510642, China

Received:2014-11-28 Revised:2015-04-03 Online:2016-02-15 Published:2016-04-26

摘要/Abstract

摘要：

苯丙氨酸解氨酶(PAL)是植物木质素生物合成途径中的关键酶,参与木质素、类黄酮和花青素等次生代谢产物的形成。本研究以‘N51’象草(Pennisetum pur pureum)为材料,通过同源克隆、3'RACE和hi-TailPCR的方法,对PAL基因进行克隆,得到PpPALcDNA序列2429bp以及DNA序列全长3907bp。PpPAL编码的蛋白包含720个氨基酸,具有典型的GTITASGDLVPLSYIAG苯丙氨酸解氨酶保守活性基序。运用生物信息学进行分析,PpPAL基因cDNA序列与玉米(Zea maysL.)ZmPAL的相似性最高为94%,与其他禾本科植物也有着较高的相似性。在氨基酸序列系统进化分析中,PpPAL与ZmPAL遗传距离最近,并与其他禾本科植物PAL聚为一类。PpPAL蛋白的三维结构模型显示,PpPAL蛋白具有典型的"海马状"结构,表明PpPAL是典型的苯丙氨酸解氨酶家族,对木质素的合成起着重要的作用。

关键词: 象草, 木质素, 苯丙氨酸解氨酶, 生物信息学

Abstract:

Phenylalanine ammonia-lyase (PAL) is a key enzyme involved in lignin biosynthesis. It catalyzes the first step of phenylpropanoid pathway, which supplies precursors for a variety of secondary metabolites including phenolics, flavonoids and anthocyanin. In this study we isolated a PAL gene from Pennisetum purpureum cv. N51 using homology cloning and RACE (Rapid Amplification of cDNA Ends) methods as well as hi-Tail PCR. The length of PpPAL cDNA sequence was 2429 bp. The nucleotide sequence of PpPAL gene was 3907 bp long and high similarity to ZmPAL, with 94% identity. The PpPAL gene encoded a 720-amino-acid protein, which contained a conserved motif of GTITASGDLVPLSYIAG. In phylogenetic tree, the PpPAL had a close genetic distance with ZmPAL, and was clustered in the same group with other gramineous plants involved in lignin biosynthesis.By predicting, the three-dimensional structure of the PpPAL protein showed a typical "sea horse" shape, which indicated that PpPAL is a typical family member of phenylalanine ammonia-lyase, should play an important role in lignin synthesis.

Key words: Pennisetum purpureum, Lignin, Phenylalanine ammonia-lyase, Bioinformatic analysis

中图分类号:

陈卫卫, 唐然, 吴昱煜, 李菲, 张向前, 解新明. 象草PAL基因克隆及其蛋白质结构与功能预测[J]. 草地学报, 2016, 24(1): 137-145.

CHEN Wei-wei, TANG Ran, WU Yu-yu, LI Fei, ZHANG Xiang-qian, XIE Xin-min. Cloning of a PAL Gene of Pennisetum purpureum and Prediction of Its Protein Structure and Function[J]. Acta Agrestia Sinica, 2016, 24(1): 137-145.

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象草PAL基因克隆及其蛋白质结构与功能预测

Cloning of a PAL Gene of Pennisetum purpureum and Prediction of Its Protein Structure and Function

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