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Detection of two exogenous genes in transgenic cattle by loop-mediated isothermal amplification

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Abstract

Nucleotide-based analytical approaches are indispensable and effective, targeting for the transgenic ingredients in biotechnical products in terms of safety assessment. In this study, a loop-mediated isothermal amplification method was developed for the specific detection of exogenous nucleic acids of hLTF/hLALBA-induced transgenic cattle. The detection limit of the LAMP method was proved to be as low as 10 copies of target molecules in optimized systems, and to be 10–100 times more sensitive than the conventional PCR. Furthermore, fluorescent dye SYBR Green I was used to visualize the color changes of LAMP products by naked eyes in daylight, which resulted in distinct colors between positive and negative reactions. For the detection of transgenes, all the transgenic samples collected from hLTF and hLALBA-induced cattle were amplified by LAMP in 1 h, followed by direct visual SYBR Green I dying or gel electrophoresis. Results showed that transgenic and non-transgenic samples exhibited distinct properties in colors or electrophoresis profiles. Thus, all the results indicated that the LAMP assay was a simple and convenient method for the test of transgenic animals.

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Acknowledgments

We are grateful to Dr. Ning Li for providing the transgenic samples. We also appreciate the help of Professor Liu Yanxiu’s help in English. This work was supported by the National Transgenic Project of China. (2009ZX08012-015B).

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Authors and Affiliations

  1. Key Lab of Agricultural Animal Genetics, Breeding, and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan, 430070, Hubei, People’s Republic of China

    Shanli Zhai, Chuxin Liu, Qingde Zhang, Chenyu Tao & Bang Liu

  2. Key Lab of Swine Genetics and Breeding of Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, Hubei, People’s Republic of China

    Shanli Zhai, Chuxin Liu, Qingde Zhang, Chenyu Tao & Bang Liu

Authors
  1. Shanli Zhai
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  2. Chuxin Liu
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  3. Qingde Zhang
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  4. Chenyu Tao
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  5. Bang Liu
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Corresponding author

Correspondence to Bang Liu.

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Zhai, S., Liu, C., Zhang, Q. et al. Detection of two exogenous genes in transgenic cattle by loop-mediated isothermal amplification. Transgenic Res 21, 1367–1373 (2012). https://doi.org/10.1007/s11248-012-9614-2

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  • DOI: https://doi.org/10.1007/s11248-012-9614-2

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