关键词: 慈竹, C3H基因, 克隆, 生物信息学分析

Abstract: p-Coumarate 3-hydroxylase plays an important role in the biosynthesis pathways of plant lignin. The full-length sequence of C3H in Neosinocalamus affinis(NaC3H) was successfully cloned by RT-PCR, and its bioinformation analysis was carried out. It aims to provide theoretical basis for decreasing the content of lignin in bamboo for making-pulp through engineering technique. The sequence is 1 581 bp in length. The bioinformation analysis showed that the CDS region of the nucleotide sequence was 1 539 bp, encoding 512 predicted amino acids. The protein molecular weight was 58.33 kD and theoretical pI was 9.09. According to the amino acid sequence and structural analysis, it showed that the protein encoding by C3H contained one conserved domain, viz. P450 domain. Phylogenetic analysis showed that NaC3H was most similar to C3H from P.edulis and Oryza sativa. The proteins encoded by C3H genes were hydrophilic ones. Subcellular localizations of these three proteins were in the endoplasmic reticulum (membrane). The secondary structures and tertiary structures were abundant in alpha helixs and random coils, beta turns and extended strains were less. Two relative conserved disordered domains were found in amino acid sequences encoded by C3H genes from N.affinis, P.edulis and O.sativa. The full-length sequence of NaC3H has been submitted to GenBank with the accession number JF693629.

Key words: Neosinocalamus affinis, C3H gene, cloning, bioinformation analysis