摘要
采用改良CTAB法从月季(Rosa hybrida)花瓣中成功提取出总RNA,进行反转录合成cDNA模板。根据GenBank已发表的其他植物CHS基因序列的保守结构域设计简并引物,采用同源克隆法进行克隆,获得RhCHS片段序列,并分析该片段的生物学信息,利用半定量RT-PCR对不同花发育时期进行表达分析。序列分析结果表明,克隆到的cDNA片段长度为860 bp,编码287个氨基酸残基,GenBank登录号KC145553。同源序列比对发现,月季RhCHS与其他植物的同源性很高,说明CHS在进化的过程中具有高度的保守性。表达谱分析表明,CHS在盛花期表达量最高。
It was to clone and identify chalcone synthase gene （ RcCHS ） from Rosa hybrida ‘Fairyland＇ , analysis the characteristic of the sequence and expression. The total RNA was isolated from the flower petal of Rosa hybrida ‘ Fairyland＇ by improved CTAB, then reverse transcription. Using PCR degenerate primers designed based on GenBank published the conserved sequences of chalcone synthase genes from other plants to amplify cDNA fragments, a new chalcone synthase gene named RcCHS was cloned. The expression patterns of RcCHS in different developmental stages were analyzed by semi-quantitative RT-PCR. The length of RcCHS was 860 bp which encoded 287 amino acids, GenBank accession No. KC145553. Homology analysis showed that the RcCHS of Rosa hybrida ‘Fairyland＇ had high homologies with other plants, and indicated that CHS in evolution process kept the high conservatism. RcCHS had a highest transcript level in petals at blooming stage.
出处
《生物技术通报》
CAS
CSCD
北大核心
2013年第7期66-70,共5页
Biotechnology Bulletin
基金
四川农业大学大学生科研兴趣培养计划经费项目(04050957)
