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Construction of Gene Library of 20 kb DNAs from Parasporal Crystal in Bacillus thuringiensis Strain 4.0718: Phylogenetic Analysis and Molecular Docking

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Abstract

The 20 kb DNAs are associated with crystals in many subspecies of Bacillus thuringiensis. We isolated 20 kb DNAs from crystals of B. thuringiensis strain 4.0718, then constructed a gene library using DNA fragments of Sau3AI partial digestion and pbluescriptIISK(+) vector. We screened out 440 recombinants, yielding a genomic coverage of ten and including 99% sequence of DNA which achieved the required theoretical value to construct the gene library. Through NCBI Blast and homology analysis, the sequencing results proved that the DNA came from the chromosome of B. thuringiensis. Moreover, we have completed the multiple alignment of homologous ropB protein sequences and phylogenetic analysis using bioinformatic software. For further investigation of the interactions between 20 kb DNAs and protoxins, molecular docking has also been done.

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Acknowledgments

This investigation was supported by National Natural Science Foundation of China (30870064, 30970066, 30900037) and National High Technology Research and Development project (863) of China (NC2010GA0091, 2011AA10A203).

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Authors and Affiliations

  1. Hunan Provincial Key Laboratory of Microbial Molecular Biology–State Key Laboratory Breeding Base of Microbial Molecular Biology, College of Life Science, Hunan Normal University, Changsha, 410081, China

    Feng Wu, Xinmin Zhao, Yunjun Sun, Wenping Li, Liqiu Xia, Xuezhi Ding, Jia Yin, Shengbiao Hu, Ziquan Yu & Ying Tang

  2. Department of Chemistry and Environmental Engineering, Hunan City University, Yiyang, 413000, Hunan, China

    Xinmin Zhao

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  1. Feng Wu
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  2. Xinmin Zhao
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  3. Yunjun Sun
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  4. Wenping Li
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  6. Xuezhi Ding
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  7. Jia Yin
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  8. Shengbiao Hu
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  10. Ying Tang
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Corresponding author

Correspondence to Liqiu Xia.

Additional information

Feng Wu and Xinmin Zhao contributed equally to this work.

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Wu, F., Zhao, X., Sun, Y. et al. Construction of Gene Library of 20 kb DNAs from Parasporal Crystal in Bacillus thuringiensis Strain 4.0718: Phylogenetic Analysis and Molecular Docking. Curr Microbiol 64, 106–111 (2012). https://doi.org/10.1007/s00284-011-0038-7

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  • DOI: https://doi.org/10.1007/s00284-011-0038-7

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