青杄中sPPa1的cDNA序列克隆及其生物信息学分析  

DNA Cloning and Bioinformatic Analysis of the sPPa1Gene from Picea wilsonii

作　　者：曹一博[1] 刘亚静[1] 张凌云[1] 

机构地区：[1]北京林业大学森林培育与保护教育部重点实验室,北京100083

出　　处：《植物科学学报》2012年第4期394-401,共8页Plant Science Journal

基　　金：国家转基因生物新品种培育科技重大专项(2009ZX08009-062B)

摘　　要：以青杄(Picea wilsonii)均一化cDNA文库为模板,通过RACE方法克隆得到青杄PPa1基因cDNA全长,对该cDNA序列、核苷酸序列的相似性、理化性质、疏水性、二级结构、三级结构及是否跨膜进行了分析预测;进行了多序列比对并构建了系统树,同时对PPa1在青杄各组织中的表达量进行了检测。结果表明:青杄PPa1基因共由216个氨基酸组成,分子量为24.55 kD,理论PI为5.83,属可溶性蛋白;二级结构主要由α-螺旋、不规则卷曲和β-折叠构成;PPa1在青杄花粉中表达量最高。研究为进一步研究青杄PPa1的功能奠定了基础。The full-length cDNA sequence of the PPal gene was obtained by the RACE method based on the cDNA library of Picea wilsonii. Bioinformatic analysis was used to predict the physicochemical properties, hydrophobicity,secondary structure,and tertiary structure of PwPPa 1. Multiple sequences alignment and phylogenetic trees were also constructed to predict the conserved domain and genetic relationship with other species. The RT-qPCR assays were used to identify the tissue expression level of PPal in Picea wilsonfi. The results showed that PwPPal consisted of 216 amino acids. The molecular weight was 24.55 kD and theoretical PI was 5.83. The PPal was a hydrophilic protein and the secondary structure of PPal was mainly composed of alpha helix,random coil,and extended strand. The expression level of PPal was highest in the pollen. This research provides the foundation for further studies on the functions of PwPPa 1.

关 键 词：青杄 PPa1 克隆 RACE 生物信息学 

分 类 号：Q78[生物学—分子生物学] Q949.665