Cloning and Expression Analysis of ClCRY2 Gene in Cunninghamia lanceolata
[Objective]By cloning the CRY2 gene of Cunninghamia lanceolata,the expression character-istics of CRY2 gene in different tissues and under different light quality treatments were studied,which provided a reference for exploring the regulatory role of CRY2 gene in the growth and development of C.lanceolata.[Method]Based on the results of our research group's full-length transcriptome sequencing of C.lanceolata,one-year old cuttings from the superior clone 061 were employed as experimental mate-rials.Using reverse transcription polymerase chain reaction(RT-PCR)technology,gene-specific prim-ers were designed and the CRY2 gene were cloned from the tender leaves of C.lanceolata seedlings,named ClCRY2(Accession number PP400318).Furthermore,bioinformatics analysis were conducted on this gene and its expression pattern were analyzed using real-time quantitative PCR technology.[Result]The bioinformatics analysis results showed that the full length of the CDs region of the ClCRY2 gene in C.lanceolata was 2 151 bp,encoding a total of 716 amino acids,with a homologous consistency of 68.93%with the AtCRY2 protein sequence in Arabidopsis.Phylogenetic tree analysis revealed that the ClCRY2 protein was most closely related to CjCRY2 in Japanese cedar(Cryptomeria japonica).The sec-ondary structure of ClCRY2 protein was mainly dominated by random coils and alpha helix,accounting for 51.26%and 34.78%,respectively.The molecular formula of ClCRY2 protein is C3 620H5 573N1 033O1 063S 27,with a theoretical isoelectric point of 6.46.The largest proportion of amino acids was leucine,accounting for 9.8%,followed by serine,accounting for 8.0%.ClCRY2 protein was a hydrophilic protein that did not have a transmembrane region and had no signal peptide.Real time quantitative PCR analysis results showed:ClCRY2 gene was observed to express in the roots,stems,and leaves of C.lanceolata seedlings,with the highest expression level in the stem,followed by the leaves,and the lowest expression level in the root.Blue light significantly increased the expression of ClCRY2 gene.[Conclusion]Through the study of the physicochemical properties and structure of the protein encoded by the ClCRY2 gene,it is speculated that this gene plays an important regulatory role in the development process of C.lanceolata stems and leaves,and participates in the conduction process of blue light signal.
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