Ganoderma lucidum, (Curtis) P. Karst. (Curtis) P. Karst.

Lu, Xiaoxiao, Xie, Chunqin, Wang, Yunxiao, Liu, Yongnan, Han, Jing, Shi, Liang, Zhu, Jing, Yu, Hanshou, Ren, Ang & Zhao, Mingwen, 2020, Function of ceramide synthases on growth, ganoderic acid biosynthesis and sphingolipid homeostasis in Ganoderma lucidum, Phytochemistry (112283) 172, pp. 1-9 : 7

5.2. Gene cloning and sequence analysis of lag in G. lucidum

The sequences of CERSs from S. cerevisiae, LAG 1 and LAC1 (National Center for Biotechnology Information (NCBI), GenBank accession number: NP_011860.1 and NP_012917.3), were used for a homology search of the genome database of G. lucidum strain 2,601,251 (NCBI taxid: 1,077,286) ( Chen et al., 2012). Three genes (lag1, lag2 and lag3 with the GenBank accession numbers MH145349 , MH145350 and MH145351 , respectively) were identified as genes encoding CERS homologues. Using the primers described in Supplementary Table S1, the three lag coding sequences were polymerase chain reaction (PCR) amplified from complementary DNA (cDNA) and sequenced. From comparisons between the cDNA and genomic sequences, the ORFs (open reading frames) and exon/intron positions of three lag s in G. lucidum were inferred.

The conserved domains were identified by SMART (http://smart. emblheidelberg.de) ( Letunic et al., 2009). The multiple sequence alignments were obtained with T-COFFEE (http://www.tcoffee.org) ( Taly et al., 2011). The phylogenetic tree was constructed through the application of MEGA 6.0 program ( Tamura et al., 2013) and the neighbour-joining method with 1000 bootstrap replicates was applied to infer evolutionary history.