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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

机译:苏云金芽孢杆菌4.0718孢子旁晶体20 kb DNA基因文库的构建:系统发育分析和分子对接

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摘要

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.
机译:苏云金芽孢杆菌许多亚种中的20kb DNA与晶体有关。我们从苏云金芽胞杆菌4.0718菌株的晶体中分离了20 kb DNA,然后使用Sau3AI部分消化的DNA片段和pbluescriptIISK(+)载体构建了一个基因库。我们筛选出440个重组体,产生的基因组覆盖率达10,包括99%的DNA序列,这些序列达到了构建基因库所需的理论值。通过NCBI Blast和同源性分析,测序结果证明该DNA来自苏云金芽孢杆菌的染色体。此外,我们已经使用生物信息学软件完成了同源ropB蛋白序列的多重比对和系统发育分析。为了进一步研究20kb DNA与毒素之间的相互作用,还进行了分子对接。

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