青杄PsbO基因cDNA序列克隆及生物信息学分析

利用RACE技术从已有的青杄均一化cDNA文库中克隆得到青杄(Picea wilsonii)PsbO基因cDNA的全长,并用生物信息学的方法对该cDNA的核苷酸序列、氨基酸序列的相似性,及编码蛋白PwPsbO的理化性质、亲水性/疏水性、二级和三级结构,PwPsbO基因进化树等进行了预测和分析.采用实时荧光定量PCR(RT-qPCR)技术,测定了青杄不同组织中的PsbO基因的相对表达水平.结果发现:青杄PsbO基因编码346个氨基酸,预测相对分子质量为36.6 kD,理论pI值为5.29,属于可溶性蛋白.二级和三级结构预测表明其含有较多的α螺旋、β折叠和随机卷曲结构.RT-qPCR发现该基因在青杄针叶和茎中表达量最高.这些结果为青杄中PsbO基因的初步功能研究奠定了基础.

cDNA Cloning and Bioinformatic Analysis of PsbO Gene from Picea wilsonii

Full-length cDNA sequence of PsbO gene was acquired from normalized cDNA library of Picea wilsonii by RACE methods.Prediction on physical and chemical properties,hydrophobicity,phylogenetic tree,secondary and tertiary structure of amino acid were analyzed by bioinformatics.The relative expression level of PwPsbO in different tissues using quantitative real time PCR(RT-qPCR) was measured.The results showed that PwPsbO gene encodes 346 amino acids.Its molecular weight is 36.6 kD and the theoretical pI is 5.29.It is a hydrophilic protein.The secondary and tertiary structure of PwPsbO is mainly composed of alpha helix,random coil and extended strand.The higher expression level of PwPsbO was observed in needles and stems.The aim of this study is to lay a foundation for the function of PsbO in Picea wilsonii.