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- Title
榅桲4CL基因的克隆与生物信息学分析.
- Authors
杨波; 车玉红; 郭春苗; 木巴热克·阿尤普; 吴津蓉; 杜鹃
- Abstract
【Objective】 To clone and study the sequence characteristics of the 4-coumarate CoA ligase (4CL) gene from the pulp of Cydonia oblonga Mill will lay a foundation of further study on the role and function of this gene in the development of quince fruit.【Methods】 The pulp of Cydonia oblonga Mill was used as test material and based on the 4CL gene cDNA sequence of related species reported by GenBank, primers were designed by primer premier 5.0 software. The total RNA was extracted and the cDNA was synthesized by reverse transcription. The 4CL gene fragment was amplified by RT-PCR and cloned into pMD 18-T vector. DNAstar software was used for homologous sequence alignment. ClustalX combined with MEGA 4.1 software was used to construct phylogenetic tree. Protparam online program was used to analyze the physical and chemical properties of proteins, and the protein program of DNAstar was used to predict the secondary structure.【Results】 The open reading frame (ORF) sequence of 4CL gene in Cydonia oblonga Mill was 1,710 BP, which encoding protein of 570 amino acid residues, with a predicted molecular mass of 68.4 kD and the highest homology with other species was 91.99%. The evolutionary relationship between the two genes was close to that of water dense peach. 4CL protein in Cydonia oblonga Mill was a hydrophilic protein, and the secondary structure of 4CL protein was mainly random coil, which was accounting for 44.27%.【Conclusion】 The full-length coding region of 4CL gene in Cydonia oblonga Mill was successfully obtained, and the structural characteristics of the sequence were confirmed.
- Publication
Xinjiang Agricultural Sciences, 2021, Vol 58, Issue 6, p1055
- ISSN
1001-4330
- Publication type
Article
- DOI
10.6048/j.issn.1001-4330.2021.06.010