马尾松PmCOMT基因的克隆及实时定量表达分析  

Cloning and Real-time Quantitative Expression Analysis on PmCOMT Gene from Pinus massoniana

作　　者：吴晓刚[1] 陈佩珍[1] 韦蔷 武星 季孔庶[1] Wu Xiaogang;Chen Peizhen;Wei Qiang;Wu Xing;Ji Kongshu(Key Laboratory of Forest Genetics and Biotechnology of Ministry of Education,Co-Innovation Center for Sustainable Forestry in Southem China,Nan- jing Forestry University,Nanjing,210037)

机构地区：[1]南京林业大学南方现代林业协同创新中心林木遗传与生物技术省部共建教育部重点试验室,南京210037

出　　处：《分子植物育种》2019年第3期789-795,共7页Molecular Plant Breeding

基　　金："十三五"国家重点研发计划项目课题(2017YFD0600304);江苏高校优势学科建设工程(PAPD);南方现代林业协同创新项目共同资助

摘　　要：在木质素生物合成里,咖啡酸-O-甲基转移酶(caffeic acid-O-methyltransferase, COMT)起重要作用。本论文以马尾松幼苗作为材料,并通过PCR技术以及RACE技术从马尾松材料中克隆出Pm COMT基因的cDNA全长。该cDNA全长为1660bp,其中预测完整ORF全长为1125bp,可编码374个氨基酸的蛋白。蛋白相对分子质量为41.81 kD;等电点(pI)为5.03,总平均亲水性为-0.311。通过生物信息学的方法对其进行分析表明,Pm COMT基因具有典型的甲基转移酶的保守结构域。通过实时荧光定量表达分析表明,COMT基因在马尾松的不同组织均有表达,其中在新梢中的表达量最高,树皮中表达量最低。Caffeic acid-O-methyltransferase (COMT)plays an important role in the lignin biosynthesis.With the seedlings of Pinus rnassoniana as materials,we used PCR and RACE to clone the full-length eDNA of PmCOMT gene from Pinus massoniana.The full length of this eDNA was 1 660 bp,the predicted full length of intact open reading frame was 1 125 bp and 374 amino acids could be encoded.The relative molecular mass of the protein was 41.81 kD,the isoeleetric point (pI)was 5.03,and the total average hydrophilicity was -0.311.Analysis by bioinformatics way showed that the PmCOMT gene had a conserved domain of typical methyltransferase.Real-time fluorescence quantitative expression analysis showed that PmCOMT gene was expressed in different tissues of Pinus massoniana,and its expression was the highest in new shoots and the lowest in bark.

关 键 词：马尾松 咖啡酸-O-甲基转移酶 基因克隆 实时荧光定量表达分析 

分 类 号：Q943.2[生物学—植物学] S791.248[农业科学—林木遗传育种]