芒果MiCOL6基因的克隆及其生物信息学和表达分析

Cloning,Bioinformatics and Expression Analysis of Mango MiCOL6 Gene

CONSTANS(CO)是光周期途径的核心调控因子,其可以整合光质和生物钟输出的信号调控植物成花。在前期转录组研究的基础上,设计基因特异性引物,通过RT-PCR扩增得到芒果的1个CO基因的cDNA全长,序列长度为678 bp,根据与拟南芥CO家族基因的相似性,将其命名为MiCOL6。生物信息学分析显示,该基因编码226个氨基酸,分子量为26.88 kDa,等电点为4.85,属于亲水性的非分泌蛋白。保守结构域和进化树分析显示,该基因仅含1个CCT结构域,属于CO基因家族的第4组;二级结构分析表明,该蛋白主要以无规卷曲和α-螺旋为主;亚细胞定位预测显示该蛋白定位在细胞质膜上的概率最大;不同花发育时期表达模式分析表明,MiCOL6基因在杧果的花芽分化期表达量最高。本研究结果为进一步研究芒果MiCOL6基因在芒果中的功能提供理论基础。

CONSTANS(CO)is a core regulator of the photoperiod pathway,which integrates light quality and clock output signals to regulate the floral transformation in plants.A full-length cDNA of MiCOL6 gene was obtained from transcriptome data analysis and then verified by RT-PCR amplification in mango.Sequences analysis showed that the open reading frame was 678 bp in length,coding 226 amino,the molecular weight and isoelectric point(pI)of MiCOL6 was 26.88 kDa and 4.85,respectively;and MiCOL6 was a non-secreted hydrophilic protein.Conserved domain and phylogenetic tree analysis showed that MiCOL6 contained only one CCT domain and belonged to the group Ⅳ of the CO gene family.The secondary structure analysis showed that the protein mainly consisted of random curl and α-helix.The subcellular localization prediction showed that MiCOL6 was likely to be localized in the cytoplasm membrane.Expression pattern analysis showed that the MiCOL6 was the highest expressed in leaves during the bud differentiation period.The results of the experiment would provide a theoretical basis for the further study on the function of MiCOL6 in mango.