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An event-specific real-time PCR detection system for the transgenic rice line 114-7-2 of producing functional human serum albumin

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Abstract

Transgenic rice 114-7-2 is a newly developed transgenic rice line of producing human serum albumin (HSA). It has attracted much attention because of its economic potential. This paper was designated to discover the integration site of the transgenic HSA rice line 114-7-2 and to establish event-specific methods for qualitative and quantitative detection of the transgenic HSA rice based on the border junction fragment. One gene fragment of 5′ flanking region was successfully isolated using the TAIL-PCR methods. The fragment sequence showed that a 454-bp junction fragment contained 75 bp of T-DNA sequence and 379 bp of rice genome DNA, which is located in chromosome 4. Event-specific real-time PCR method for HSA rice line 114-7-2 was established with the primers (HSA-F/HSA-R) and the probe (HSA-P) targeting the 454-bp junction region. The qualitative PCR assay showed the limit of detection was 0.01 %. In the event-specific quantitative detection method, the LOQ for 114-7-2 HSA rice was estimated to be 0.025 ng or 50 copies. The method developed in this study is highly specific, sensitive, and reliable for transgenic HSA rice sample detection.

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Acknowledgments

We thank Daichang Yang (College of Life Sciences, Wuhan University) for providing HSA rice samples and suggestions on the manuscript. We are grateful for the support of the Important National Science & Technology Specific Projects (2013ZX08012-001) and AQSIQ Non-profit Industry Research Projects (20141004).

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None.

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This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. Institute of Plant Quarantine, Chinese Academy of Inspection and Quarantine, Beijing, 100029, China

    Lili Zhang, Xin Huang & Shuifang Zhu

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  1. Lili Zhang
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  2. Xin Huang
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  3. Shuifang Zhu
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Corresponding authors

Correspondence to Xin Huang or Shuifang Zhu.

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Zhang, L., Huang, X. & Zhu, S. An event-specific real-time PCR detection system for the transgenic rice line 114-7-2 of producing functional human serum albumin. Eur Food Res Technol 239, 403–408 (2014). https://doi.org/10.1007/s00217-014-2234-8

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  • DOI: https://doi.org/10.1007/s00217-014-2234-8

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