| 粗毛栓菌诱变菌株SAH-12漆酶基因的克隆与序列分析 |
| Cloning and sequence analysis of a laccase gene from mutated strain SAH-12 of Trametes gallica |
| 投稿时间：2006-07-30 修订日期：2006-10-17 |
| 中文关键词:cDNA RACE 分子克隆 |
| 英文关键词:cDNA RACE molecular cloning |
| 基金项目:四川省教育厅重点科研项目(2002A009,2004A026);四川省科学技术厅应用基础项目(04JY029-032) |
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| 中文摘要: |
| 粗毛栓菌Trametes gallica诱变菌株SAH-12是通过紫外诱变选育得到的漆酶高产菌株。为了对其漆酶基因进行研究和利用,采用cDNA末端快速扩增(Rapid Amplification of cDNA Ends,RACE)技术,从T.gallica诱变菌株SAH-12分离得到漆酶基因全长cDNA Lacc1(GenBank accession No.DQ431716)及其对应的结构基因Lac1(DQ431715)。该基因属于真菌漆酶基因家族,与来自出发菌T.gallica漆酶基因lacA(AY875867)在成熟肽编码区的同源性最高(一致性为98%)。Lacc1全长1891bp,由40bp的5'-UTR、1554bp的完整ORF和297bp的3'-UTR构成,具有polyA加尾信号AATACA和59bp的polyA结构;其完整ORF可编码21个氨基酸残基组成的信号肽和496个氨基酸残基组成的成熟蛋白。在Lacc1基因的推导氨基酸序列中有4个潜在的N-糖基化位点和4个参与二硫键形成的Cys残基,且含有真菌漆酶Ⅰ、Ⅱ、Ⅲ型铜离子结合区的4个高度保守序列。结构基因Lac1全长2338bp,含10个内含子和11个外显子，各内含子长度在51bp~76bp之间，且其序列均符合5'-gt…ag-3'规则。 |
| 英文摘要: |
| Laccase overproducing Trametes gallica strain SAH-12 was obtained from parent strain Trametes gallica by UV mutagenesis. Using RACE (rapid amplification of cDNA ends) method, a full-length cDNA of laccase gene and its corresponding genomic sequence, designated as Lacc1 (DQ431716) and Lac1 (DQ431715), were cloned from mutated strain SAH-12 of Trametes gallica, respectively. Sequencing analysis suggested that this gene belongs to fungi laccase gene family, and there are 98% homology in the nucleotide sequence and the deduced amino acid sequence of Lacc1 and the lacA gene from original strain T.gallica. The full length of Lacc1 was 1891bp,with 40,1554 and 297bp of 5' untranslated region,open reading frame (ORF) and 3'untranslated region,respectively.It contains polyadenylation signal AATACA and 59bp PolyA.The ORF of Lacc1 encode a polypeptide comprised of 517 amino acid residues,which included a signal peptide of 21 amino acid residues and a mature protein of 496 amino acid residues,which included a signal peptide of 21 amino acid residues and a mature protein of 496 amino acid residues.In the deduced amino acid sequence of Lacc1 gene,there are 4 putative N-glycosylation sites,4 Cys residues for forming disulfide bonds,and also 4 hihgly conserved amino acid sequences,which could combine with Ⅰ,Ⅱ and Ⅲ type copper.The full length of Lac1 gene was 2338bp,with 10 introns and 11 exons.Introns ranged in size from 51 to 76bp,and ist sequence complied with the rule of“5'-gt”and“ag-3'”. |
| 黄乾明,谢君,杨婉身. 粗毛栓菌诱变菌株SAH-12漆酶基因的克隆与序列分析[J]. 菌物学报, 2007, 26(1): 97-105 |
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