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Detection and Identification of Vegetative Insecticidal Proteins vip3 Genes of Bacillus thuringiensis Strains Using Polymerase Chain Reaction-High Resolution Melt Analysis

Current Microbiology volume 64, pages 463–468 (2012)Cite this article

Abstract

In this study, vegetative insecticidal proteins vip3 genes from Bacillus thuringiensis strains were detected based on polymerase chain reaction–high resolution melt (PCR–HRM) analysis. A pair of primers was designed according to the conservative sequences in 150 bp region of the known vip3 subfamily. The 150 bp regions of difference vip3 genes have only a few nucleotide difference vip3 genes were detected in 8 of 11 standard B. thuringiensis strains, and vip3Aa genes, vip3Af genes and vip3Ba gene can be distinguished as different melting curves by this method. The results demonstrate the utility of the HRM assay for mutant screening using vip3 gene. The PCR–HRM method may be a valuable and reliable tool for specific detection and identification of vip3 genes.

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Acknowledgments

We are grateful to professor Jie Zhang and Fuping Song (Institute of Plant Protection, Chinese Academy of Agricultural Sciences) for the help of this experiment. This study was supported by the national high technology research and development program subject (2011aa10a203), the national new varieties of genetically modified organism special science and technology major (No. 2009ZX08009-030B) and soybean biology ministry of education key laboratory open subject.

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Affiliations

  1. Northeast Agricultural University, Harbin, 150030, People’s Republic of China

    Haitao Li, Changlong Shu, Xiaoming He, JiGuo Gao & Rongmei Liu

  2. State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100094, People’s Republic of China

    Changlong Shu

  3. Institute of Biotechnology Research, Chinese Academy of Agricultural Sciences, Beijing, 100081, People’s Republic of China

    Dafang Huang

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  1. Haitao Li
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  2. Changlong Shu
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  3. Xiaoming He
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  4. JiGuo Gao
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  5. Rongmei Liu
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  6. Dafang Huang
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Correspondence to Haitao Li.

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Changlong Shu and Xiaoming He contributed equally to this work.

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Li, H., Shu, C., He, X. et al. Detection and Identification of Vegetative Insecticidal Proteins vip3 Genes of Bacillus thuringiensis Strains Using Polymerase Chain Reaction-High Resolution Melt Analysis. Curr Microbiol 64, 463–468 (2012). https://doi.org/10.1007/s00284-012-0092-9

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  • DOI: https://doi.org/10.1007/s00284-012-0092-9

Keywords

  • Melting Curf
  • High Resolution Melting
  • Insecticidal Protein
  • High Resolution Melting Analysis
  • Direct Sequencing Method