黄秋葵查尔酮合成酶基因AeCHS的克隆与表达分析

采用同源序列克隆和RT-PCR技术,首次克隆得到黄秋葵查尔酮合成酶基因(CHS)cDNA全长序列。序列分析表明,该序列全长1175 bp,包括一个1170 bp的完整ORF,编码389个氨基酸,命名为AeCHS。生物信息学分析表明,本研究所获得的AeCHS氨基酸序列与同科植物黄蜀葵和陆地棉的同源性较高,分别达99.23%和97.44%,AeCHS推断的氨基酸序列含有CHS蛋白的标签序列GFGPG以及4个保守活性位点Cys164、Phe215、His303、Asn336。实时荧光定量PCR分析黄秋葵果实、花、叶片不同发育时期AeCHS基因的表达量,结果表明AeCHS基因在上述植物材料中表现出不同的表达模式:花>果实>叶片,具体到不同植物组织,AeCHS基因在生长6 d的果实、盛开的花朵以及植株顶端第4片叶子中的表达量较高。

loning and expression analysis of chalcone synthase gene(AeCHS) in Abelmoschus esculentus L.

The full length cDNA sequence of chalcone synthase gene (CHS) from Abelmoschus esculentus was isolated by homologous sequences cloning and RT-PCR. This sequence is 1175bp, and it predicts an open reading frame of 1170bp corresponding to 389 amino acids, which is designated as AeCHS. Bioinformatics analysis shows that the deduced AeCHS amino acid sequence has the higher homology with Abelmoschus manihot and Gossypium hirsutum, respectively 99.23% and 97.44%. The deduced AeCHS amino acid sequence contains a chalcone synthase family tag sequence(GFGPG) and four conserved active sites: Cys164, Phe215, His303, Asn336. The expression analysis of AeCHS gene in different growth period of okra fruits, flowers, leaves are carried out by RT-qPCR and the results indicate that AeCHS gene shows different expression patterns: flower > fruit > leaf. Specific to different plant tissues, the expression of AeCHS gene reachs a higher level in fruits grown six days, blooming flowers and the fourth leaves numbering from the top of plants.