Engineering strain of Brasil diazotrophic spirillum DraT* containing multiple copied nifA gene
A Brazilian Azospirillum, engineering strain technology, applied in bacteria, organic fertilizers and other directions
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[0017] The present invention will be further described through construction examples and application examples of the engineering strain YuMA below.
[0018] 1. Construction of engineering strain YuMA
[0019] (1) Azospirillum brasiliensis DraT - Mutant strain construction
[0020] In the present invention, the total chromosomal DNA is first extracted from the genome library of Azospirillum brasiliensis Yu62, then digested with Sau3A, and a DNA fragment of 8-20 kb is recovered, connected to the phage carrier EMBL3A (Huamei Bioengineering Co., Ltd.) and packaged in vitro. A gene library of Azospirillum brasiliensis Yu62 was constructed. Part of the draT gene fragment of the Azospirillum brasiliensis Sp7 strain was used as a probe to screen a positive clone from the Azospirillum brasiliensis Yu62 gene library, and the length of the positive clone was 8.0 kb. The positive clone was digested with SalI, and the 8.0 kb foreign fragment containing the draT gene was recovered and clon...
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