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Article

Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation

  • Authors:
    • Jing Qi
    • Zhen Dong
    • Yu‑Xing Zhang
  • View Affiliations / Copyright

    Affiliations: College of Biology Science and Engineering, Hebei University of Economics and Business, Shijiazhuang, Hebei 050061, P.R. China, Hebei Women's Vocational College, Shijiazhuang, Hebei 050091, P.R. China, College of Horticulture, Agriculture University of Hebei, Baoding, Hebei 071001, P.R. China
  • Pages: 8268-8274
    |
    Published online on: October 7, 2015
       https://doi.org/10.3892/mmr.2015.4419
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Abstract

The aim of the present study was to genetically modify plantlets of the Chinese yali pear to reduce their expression of ripening-associated 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) and therefore increase the shelf-life of the fruit. Primers were designed with selectivity for the conserved regions of published ACO gene sequences, and yali complementary DNA (cDNA) cloning was performed by reverse transcription quantitative polymerase chain reaction (PCR). The obtained cDNA fragment contained 831 base pairs, encoding 276 amino acid residues, and shared no less than 94% nucleotide sequence identity with other published ACO genes. The cDNA fragment was inversely inserted into a pBI121 expression vector, between the cauliflower mosaic virus 35S promoter and the nopaline synthase terminator, in order to construct the anti‑sense expression vector of the ACO gene; it was transfected into cultured yali plants using Agrobacterium LBA4404. Four independent transgenic lines of pear plantlets were obtained and validated by PCR analysis. A Southern blot assay revealed that there were three transgenic lines containing a single copy of exogenous gene and one line with double copies. The present study provided germplasm resources for the cultivation of novel storage varieties of pears, therefore providing a reference for further applications of anti‑sense RNA technology in the genetic improvement of pears and other fruit.
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Copy and paste a formatted citation
Spandidos Publications style
Qi J, Dong Z and Zhang YX: Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation. Mol Med Rep 12: 8268-8274, 2015.
APA
Qi, J., Dong, Z., & Zhang, Y. (2015). Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation. Molecular Medicine Reports, 12, 8268-8274. https://doi.org/10.3892/mmr.2015.4419
MLA
Qi, J., Dong, Z., Zhang, Y."Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation". Molecular Medicine Reports 12.6 (2015): 8268-8274.
Chicago
Qi, J., Dong, Z., Zhang, Y."Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation". Molecular Medicine Reports 12, no. 6 (2015): 8268-8274. https://doi.org/10.3892/mmr.2015.4419
Copy and paste a formatted citation
x
Spandidos Publications style
Qi J, Dong Z and Zhang YX: Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation. Mol Med Rep 12: 8268-8274, 2015.
APA
Qi, J., Dong, Z., & Zhang, Y. (2015). Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation. Molecular Medicine Reports, 12, 8268-8274. https://doi.org/10.3892/mmr.2015.4419
MLA
Qi, J., Dong, Z., Zhang, Y."Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation". Molecular Medicine Reports 12.6 (2015): 8268-8274.
Chicago
Qi, J., Dong, Z., Zhang, Y."Complementary DNA cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation". Molecular Medicine Reports 12, no. 6 (2015): 8268-8274. https://doi.org/10.3892/mmr.2015.4419
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