利用SSH技术鉴定香蕉体细胞胚发生相关基因

本研究以野生蕉胚性悬浮系为材料,将胚性细胞增殖培养基上的香焦胚性悬浮系作为对照,体胚诱导培养基上的培养材料为处理,利用抑制性消减杂交(suppression subtractive hybridization,SSH)技术,构建野生蕉胚性悬浮系激素诱导的胚胎发育差异表达cDNA文库,随机挑选阳性克隆并进行生物信息学分析...

Identification of Somatic Embryo Development-Related Genes in Banana Using SSH

A subtracted cDNA library of was constructed with the embryogenic cell suspensions (ECS) of the wild banana by using Suppression Subtractive Hybridzation (SSH). The ECS subcultured in cell multiplication medium were harvested as control materials, and the ECS subcultured in embryo development medium were harvested as the treated materials. Positive clones were randomly selected and sequenced from the SSH-cDNA library. After the removal of duplication and sequencing of low-quality sequences, 62 high quality sequences were obtained, According to BLAST screening and functional annotation, the results revealed that 46 ESTs have high homology with known genes in GenBank, 12 ESTs homologous sequence were not found, presumably may be new genes. The differentially expressed genes are involved in primary metabolism, protein binding, transport, cell differentiation and et al. Among these ESTs, TPD1, heat shock proteins, and zinc finger protein and cell wall hydroxyproline-rich glycoprotein are probably related to embryo development of banana. The SSH cDNA library can be further used for gene cloning and identification for the development-related genes in banana.