摘要
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2 165 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1 873 bp,其中包含一个完整的ORF,长度为1 563 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.
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 , two elements of STRE, one element of HSEs and seven nitrogen factor binding sites , etc.The laccase gene expression of the Lenzites gibbosa can be regulated by different exterior inducers .
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2013年第4期524-530,共7页
Journal of South China Agricultural University
基金
国家自然科学基金(30671700)
中国博士后科学基金(20090460866)
齐齐哈尔市科技局社会发展攻关项目(SF-GG-201204)
关键词
偏肿革裥菌
漆酶基因
启动子克隆
转录调控
Lenzites gibbosa
laccase gene
promoter cloning
transcriptional regulation
