Cloning and sequence analysis of laccase gene lac1 from Colletotrichum gloeosporioides on mango.
Wei YunXie, Liu XiaoMei, Zhang He, Zhang Xin, Qi YanXiang, Xie YiXian, Lu Ying, Cao ShenWen, Pu JinJi
Author Affiliation: College of Environment and Plant Protection, Hainan University, Key Laboratory of Protection and Development Utilization of Tropical Crop Germplasm Resources (Hainan University), Ministry of Education, Haikou, Hainan 570228, China.
Journal of Fruit Science 30 : 202-206
Abstract : Objective: The objective of this study was to reveal sequence features of laccase(lac1), which would be used in further study of pathogenesis of C. gloeosporioides on Mango. Method: A laccase gene homolog lac1 was cloned from C. gloeosporioides on mango by the method of homologous cloning, SEFA-PCR and RT-PCR, and sequence characterization and the conserved domains on predicted protein of lac1 gene were also analyzed. Result: The results showed that the complete DNA and cDNA sequence of lac1 gene were 2 996 bp and 1 773 bp in size, respectively, with three introns of 47 bp, 50 bp and 59 bp, respectively, encoding a putative protein of 590-amino acid with a molecular weight of 1174.50 kDa and isoelectric point of 5.36. Phylogenetic analysis indicated that putative protein of lac1 was homologous to C. lagenarium laccase (BAB32575.1) with a similarity of 73%, and was also homologous to copper-containing oxidases in some other fungi at some similarities. Conclusion: The studies suggested that lac1 gene had the sequence characteristics of laccase gene family in fungi, which meant that LAC1 might involve in mycelia growth, pigment production and pathogenicity of C. gloeosporioides.