赤桉CCoAOMT亚家族的基因克隆及可变剪接分析

doi:10.11707/j.1001-7488.20140508

Abstract

Abstract:

In this study, the homology-based RT-PCR method and the rapid amplification of cDNA ends (RACE) method were used to clone full-length cDNAs of two CCoAOMT subfamily genes in Eucalyptus camaldulensis. The full-length cDNA of CCoAOMT 1 gene is 1 020 bp and contains 738 bp ORF, 106 bp 5'-UTR and 176 bp 3'-UTR, and the full-length cDNA of CCoAOMT 2 gene is 1 047 bp and contains 741 bp ORF, 125 bp 5'-UTR and 158 bp 3'-UTR.The alignment result showed that CCoAOMT 1 and CCoAOMT 2 genomic DNA both contain five exons. Their main difference lay in that there was a alternative 5' splice site at the CCoAOMT 1 's first intron, which could result in formation of two mRNA with different lengths. One mRNA contains 738 bp ORF, and the other has a 42 bp deletion, however this splicing doesn't change the reading frame sequence of posterior amino acids. The alternative splicing of CCoAOMT 1 gene expresses from May to September, at the most vigorous growing stages of Eucalyptus stem, suggesting that this alternative splicing may involve in the development or growth of tissues and organs. Analysis result of protein's three-dimensional structure showed that the CCoAOMT can form the correct active center of enzyme and the substrate binding site. Limitation of the CCoAOMT genes' expression by genetic manipulation may serve as a way to reduce the lignin content in Eucalyptus, which presents a significant prospect in reducing pollutants generated from pulping industry.

Key words: Eucalyptus, lignin, CCoAOMT subfamily, gene cloning, alternative splicing, bioinformatics annotation

CLC Number:

S718.46

Gu Zhenjun, Zhang Huaiyun, Zhang Dangquan, Xie Yaojian, He Hanjie, Chen Lili, Peng Kuan, Liu Guo, Yang Dan. Gene Cloning and Alternative Splicing of CCoAOMT Subfamily Genes from Eucalyptus camaldulensis[J]. Scientia Silvae Sinicae, 2014, 50(5): 62-69.

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Gene Cloning and Alternative Splicing of CCoAOMT Subfamily Genes from Eucalyptus camaldulensis

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