偏肿革裥菌漆酶基因克隆及启动子序列分析

以优化后的漆酶培养基为基础，通过RT-PCR和RACE技术相结合，从偏肿革裥菌Lenzites gibbosa菌株中获得编码漆酶基因的cDNA及Genomic DNA的全长序列，Genomic DNA大小为2165 bp．通过比较该漆酶基因的cDNA和Genomic DNA的全长序列，发现该基因包含11个外显子和10个内含子．cDNA序列的全长为1873 bp，其中包含一个完整的ORF，长度为1563 bp，编码520个氨基酸．序列在氨基酸水平上与彩绒革盖菌Trametes versicolor的相似性评价最高，相似性达83％．通过SEFA-PCR的方法，扩增得到漆酶基因起始密码子上游长986...

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Bibliographic Details
Published in华南农业大学学报 Vol. 34; no. 4; pp. 524 - 530
Main Author 郑苗苗 池玉杰
Format Journal Article
LanguageChinese
Published 齐齐哈尔大学生命科学与农林学院，黑龙江齐齐哈尔161006 2013
东北林业大学林学院，黑龙江哈尔滨150040%东北林业大学林学院,黑龙江哈尔滨,150040
Subjects
偏肿革裥菌
启动子克隆
漆酶基因
转录调控
Lenzites gibbosa
偏肿革裥菌
漆酶基因
启动子克隆
promoter cloning
转录调控
laccase gene
transcriptional regulation
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ISSN1001-411X

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Summary:以优化后的漆酶培养基为基础，通过RT-PCR和RACE技术相结合，从偏肿革裥菌Lenzites gibbosa菌株中获得编码漆酶基因的cDNA及Genomic DNA的全长序列，Genomic DNA大小为2165 bp．通过比较该漆酶基因的cDNA和Genomic DNA的全长序列，发现该基因包含11个外显子和10个内含子．cDNA序列的全长为1873 bp，其中包含一个完整的ORF，长度为1563 bp，编码520个氨基酸．序列在氨基酸水平上与彩绒革盖菌Trametes versicolor的相似性评价最高，相似性达83％．通过SEFA-PCR的方法，扩增得到漆酶基因起始密码子上游长986 bp的启动子序列．分析表明，该启动子区域上除分布有TATA-box、CAAT-box以及AP2等基本的转录起始元件外，还存在有多个潜在的顺式作用元件序列位点，包括7个MRE元件、2个STRE元件、1个HSEs元件、7个氮因子结合位点等．这些结果表明，不同的外源诱导物可以调节偏肿革裥菌漆酶基因的表达．
Bibliography:Lenzites gibbosa ; laccase gene ; promoter cloning; transcriptional regulation
ZHENG Miaomiao, CHI Yujie （1 College of Life Science and Agriculture Forestry, Qiqihar University, Qiqihar 161006, China; 2 College of Forestry, Northeast Forestry University, Harbin 150040, China）
44-1110/S
The cDNA and Genomic DNA sequences of the laccase gene from Lenzites gibbosa were ob-tained by PCR and RACE technology , and the length of laccase gene was 2 165 bp.Comparison of the cDNA and DNA sequences showed that the laccase gene contained 11 exons and 10 introns.The cDNA length of laccase gene was 1 873 bp, including an ORF with 1 563 bp length and codes 520 amino acids . The closest organism was Trametes versicolor with 83%similarity level of amino acids .The 986 bp se-quences of promoter located in the upstream of start code of laccase gene were obtained by the approach of SEFA-PCR.The promoter not only scattered in the basic transcriptional elements of TATA-box, CAAT-box and AP2, but also contained seven elements of MRE ,
ISSN:1001-411X