1. 樟叶越桔肌动蛋白 Actin7 基因克隆与组织表达.
- Author
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蒲芝雨, 杨玉菊, 张安勉, 赵平, and 丁勇
- Subjects
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GENE expression , *TRANSMEMBRANE domains , *MOLECULAR cloning , *ISOELECTRIC point , *LEAF development , *GENETIC code - Abstract
【Objective】The present paper aimed to study VdActin7 gene expression pattern in different tissues of Vaccinium dunalianum Wight.【Method】The cDNA and DNA of VdActin7 gene were successfully cloned by PCR technique based on the transcriptome database of 3 generations of V. dunalianum. The expression of VdActin7 gene in young leaves, mature leaves, flower buds, flowers, green fruits, red fruits, green fruit stems and red fruit stems of V. dunalianum was further detected and analyzed using qRT-PCR technique. 【Result】The full length of VdActin7 gene and cDNA were 1657 and 1387 bp, respectively, which contained 3 introns, namely, intron1 of 93 bp, intron2 of 89 bp and intron3 of 88 bp. It had an open reading frame length of 1134 bp, encoding 377 amino acids. The protein VdActin7 with a relative molecular weight of 41.70 KD and a theoretical isoelectric point of 5.31, was a stable hydrophilic protein, without signal peptide and transmembrane domain. Phylogenetic analysis revealed that the VdActin7 was closest to V. ashei Actin7, which also belonged to the Vaccinium family. The codon usage bias analysis of the VdActin7 gene showed that GC contents of codon(GC) and 3rd nucleotide in codon GC3s were 48.2% and 44.1%, respectively, the effective number of codon(ENC) was 49.28%, a total of high-frequency codons was 31. The codon bias was similar to that of Ziziphus, both with the strongest bias for CUU, followed by GUU, and the bias for CUU was significantly stronger in V. dunalianum(3.43) than that in Ziziphus(2.57). qRT-PCR analysis showed that the VdActin7 gene expression showed obvious tissue specificity, with the highest in young leaves while lower expression in mature leaves and flowers, meanwhile no detectable expression or no expression in fruits, suggesting that the VdActin7 gene might play an important role in the development of young leaves. 【Conclusion】In the paper, we reported the Actin7 gene sequence and its characteristics in V. dunalianum for the first time, and found that the VdActin7 gene had tissue expression specificity. This laid the foundation for further study of VdActin7 gene function, expression regulation and mechanism of action. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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