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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。
The full-length cDNA of chalcone synthase gene (CHS) was isolated from the first clone by homologous sequence cloning and RT-PCR. Sequence analysis showed that the full length of this sequence was 1175 bp, including a complete 1170 bp ORF encoding 389 amino acids and named AeCHS. Bioinformatics analysis showed that the amino acid sequence of AeCHS obtained in this study has high homology of 99.23% and 97.44% with the plants of Hollyhock and Upland cotton, AeCHS deduced amino acid sequence of the gene sequence of GFGPG containing CHS protein and Four conserved active sites Cys164, Phe215, His303, Asn336. The results showed that the AeCHS gene showed different expression patterns in the above plant materials: flower> fruit> leaf, specific to different plant tissues, AeCHS gene The fruit of the 6th day of growth, the flower in full bloom, and the 4th leaf at the top of the plant expressed higher level.