CODEHOP在线简并引物设计与杉木4CL基因片段克隆  

Degenerate primer online design using CODEHOP software and the clone of 4-CL gene fragment of Cunninghamia lanceolata

作　　者：佘朝文[1,2,3] 蒋向辉[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：《湖南林业科技》2009年第6期26-29,共4页Hunan Forestry Science & 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4CL酶的简并引物,利用设计的3对简并引物进行RT—PCR,从杉木幼茎中克隆到2个长度分别为502 bp和536 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Blastx检索与Genbank进行同源性比对后,发现2片段都与其它植物的4—CL基因具有较高的氨基酸序列同源性。研究表明,该程序设计的简并引物可信性强,阳性率高。该基因的成功克隆将为研究杉木木质素的合成提供科学依据。This work used CODEHOP to online design the degenerate primers of 4-CL enzyme, chose the designed three pairs of degenerate primers to carry out RT-PCR. 2 cDNA fragments with the length of 502 bp and 536 bp respectively were cloned from young stem of Curmingharnia lanceolata, which were transformed into the E. coli DH5α through being/inked with pMD18-T vector and sequenced after filtration. Similarity alignment showed that the products of the cloned DNA were very similar to those of 4-CL gene from other species. The cloned sequence was putatively 4-CL gene DNA fragment. The results indicated that these degenerate primers designed by the CODEHOP software could be used to obtain specific gene fragment with strong credibility and high positive rate. The success of cloning this gene fragment would provide scientific thereunder for research on lignin biosynthesis of Cunninghamia lanceolata.

关 键 词：CODEHOP 简并引物 4-CL基因 克隆 

分 类 号：S791.27[农业科学—林木遗传育种]