Cloning and expression analysis of FabHLH3 gene from cultivated strawberry and construction of overexpression and silencing vector

doi:10.3969/j.issn.1004-1524.2017.02.07

›› 2017, Vol. 29 ›› Issue (2): 220-227.DOI: 10.3969/j.issn.1004-1524.2017.02.07

• Horticultural Science • Previous Articles Next Articles

Cloning and expression analysis of FabHLH3 gene from cultivated strawberry and construction of overexpression and silencing vector

ZHANG Qing¹, MIAO Lixiang², ZHANG Yuchao², YANG Xiaofang², JIANG Guihua^{2, *}

1.Agricultural Technology Extension Centre of Lin'an, Lin'an 311300, China;;
2.Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2016-10-14 Online:2017-02-15 Published:2017-03-06

Abstract

Abstract: The full-length cDNA sequence of FabHLH3 gene was cloned from strawberry (Fragaria×ananassa cv. Yuexin) through homology cloning method. Then, bioinformatics and expression pattern of FabHLH3 gene were analyzed by some softwares and real-time fluorescent quantitative PCR technique, respectively. Sequence analysis showed that the cDNA of FabHLH3 was 2 094 bp, encoding 697 amino acids, while the estimated molecular weight and isoelectric point of the putative protein were 77.43 ku and 6.05. After DNAMAN analysis, it shared 92.04% and 90.87% identity with the nucleotide sequence and amino acid sequence of bHLH3 gene from woodland strawberry(F. vesca), respectively. The expression of FabHLH3 was analyzed in leaf, petiole, flower and fruit being grown under colored light-quality plastic film. The results showed that FabHLH3 was expressed in all the tissues and the highest level was found in leaf. The expression analysis showed that the expression level of FabHLH3 was gradually increased during fruit development. The lowest expression was detected at small green fruit stage, and a reduced peak appeared at the full red stage. In addition, the expression of FabHLH3 gene can be induced by red and blue light. The overexpression and silencing vectors of FabHLH3, pGreen-FabHLH3 and pGreen-FabHLH3i, were constructed by in-fusion method with a plant expression vector pGreenII 62-SK. It would be used in functional analysis in the future research.

Key words: strawberry, FabHLH3, gene cloning, expression pattern, overexpression and silencing vector

CLC Number:

S668.4

ZHANG Qing, MIAO Lixiang, ZHANG Yuchao, YANG Xiaofang, JIANG Guihua. Cloning and expression analysis of FabHLH3 gene from cultivated strawberry and construction of overexpression and silencing vector[J]. , 2017, 29(2): 220-227.

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Cloning and expression analysis of FabHLH3 gene from cultivated strawberry and construction of overexpression and silencing vector

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