Liqiu Xia
Key Laboratory of Microbial Molecular Biology of Hunan Province, Life Science College of Hunan Normal University, Changsha 410081, ChinaShengbiao Hu
(1Key Laboratory of Microbial Molecular Biology of Hunan Province, Life Science College of Hunan Normal University, Changsha 410081, China;2State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China)Li Yan
Key Laboratory of Microbial Molecular Biology of Hunan Province, Life Science College of Hunan Normal University, Changsha 410081, ChinaXuezhi Ding
Key Laboratory of Microbial Molecular Biology of Hunan Province, Life Science College of Hunan Normal University, Changsha 410081, ChinaYouming Zhang
Key Laboratory of Microbial Molecular Biology of Hunan Province, Life Science College of Hunan Normal University, Changsha 410081, ChinaZiniu Yu
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, ChinaSupported by the National Natural Science Foundation of China (30570050, 30670052) ,the National Programs for High Technology Research and Development of China (2006AA02Z187, 2006AA10A212) and the Research Fund for the Doctoral Program of Higher Education
[Objective] To efficiently construct resistance gene-free Bacillius thuringiensis engineered strain that can stably express heterologous gene. [Methods] We amplified the trigger factor gene located in chromosome of XBU001 strain as homologous arms and constructed an integrative plasmid pKTF12 on the basis of plasmid pKSV7, a temperature sensitive plasmid. We also constructed a recombinant strain KCTF12 containing cry1Ac gene in its chromosome via the integrative plasmid pKTF12. [Results] Site-specific integration of cry1Ac into XBU001 chromosome did not affect its normal growth. The cry1Ac gene could stably express and form bipyramid crystals in KCTF12. When compared with HTX42 harboring a high-copy number plasmid, the recombinant strain KCTF12 has the merit of advanced sporulation and an increase in spore number. [Conclusion] The Site-specific integration proved to be a good approach to construct resistance gene-free Bacillius thuringiensis engineered strain that can stably express the heterologous gene.
Ping Liu, Liqiu Xia, Shengbiao Hu, Li Yan, Xuezhi Ding, Youming Zhang, Ziniu Yu. Site-specific integration of heterologous gene into Bacillus thuringiensis chromosome and its expression. [J]. Acta Microbiologica Sinica, 2008, 48(5): 661-666
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