杉木CCoAOMT基因部分cDNA克隆与序列分析  

Partial cDNA Cloning and Sequence Analysis of CCoAOMT Gene of Cunninghamia lanceolata(Lamb.)

作　　者：蒋向辉[1,2,3] 佘朝文[1,2,3] 许栋[1] 张青桦[1] 赵旺[1] 

机构地区：[1]怀化学院生命科学系,湖南怀化418008 [2]怀化学院民族药用植物资源研究与利用湖南省重点实验室,湖南怀化418008 [3]怀化学院湘西药用植物与民族植物学湖南省高校重点实验室,湖南怀化418008

出　　处：《中南林业科技大学学报》2009年第6期24-28,共5页Journal of Central South University of Forestry & Technology

基　　金：湖南省教育厅重点资助项目(06A054);湖南省科研计划重点资助项目(2009FJ2008);民族药用植物资源研究与利用湖南省重点实验室项目(SYSXM200909)

摘　　要：利用CODEHOP设计CCoAOMT基因的简并引物,通过RT-PCR的方法,从杉木茎段中克隆到1个长度为614 bp的PCR产物,产物经pMD18-T载体克隆转化至DH5α大肠杆菌中,序列通过同源性比对后显示该片段与其它植物的CCoAOMT基因具有较高的同源性.利用生物信息学方法分析发现,该序列在112－651 bp片段上有一个开放性阅读框,该序列编码的是一个酸性蛋白,亲水性较弱,疏水性较强;信号肽序列为第23位至第45位,二级结构以α-螺旋（40.62%）与不规则盘绕（45.31%）为主,没有发现β转角区域.This work used CODEHOP to make online design of degenerate primers of CCoAOMT gene.A cDNA fragment of 614 bp was cloned from Cunninghamia lanceolata（Lamb.） Hook.,which was transformed into the E.coli DH5α through being ligated with pMD18-T vector and sequenced after filtration.Similarity alignment shows that the product of the cloned DNA has high homology with CCoAOMT gene and the cloned sequence is putatively CCoAOMT gene cDNA fragment.Using a series of bioinformatics software analysis,the results show that the sequence contains one ORF during 112651 bp,the sequence codes an acidic protein which has features of less hydrophilic,more hydrophobicity,and signal peptide located in section 23 to section 45,in the structure α-helix and random coil are 40.62% and 45.31% respectively,β-angle region isn＇t found,these are main features of protein secondary structure that was predicted.

关 键 词：生物技术 杉木 CCoAOMT基因 CDNA克隆 序列分析 

分 类 号：Q785[生物学—分子生物学]