Nocardia seriolae attenuated strain and preparation method and application thereof
A technology of Nocardia amberjack and an attenuated strain is applied in the field of genetic engineering, and can solve the problems of antibiotic resistance, accumulated poisoning in the human body, and long treatment course of treatment, and achieves a short cycle, reduced pathogenicity, and improved preparation. simple method effect
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Embodiment 1
[0027] This embodiment provides an attenuated strain of Nocardia japonica, which is a Nocardia japonica strain containing a deletion of the alanine dehydrogenase gene; the nucleotide of the alanine dehydrogenase gene The sequence is shown in the sequence listing SEQ ID NO: 1, the amino acid sequence of the alanine dehydrogenase is shown in the sequence listing SEQ ID NO: 12, wherein the nucleotides of the upstream fragment of the alanine dehydrogenase gene The sequence is shown in SEQ ID NO:2 in the sequence listing, and the nucleotide sequence of the downstream fragment of the alanine dehydrogenase gene is shown in SEQ ID NO:3 in the sequence listing.
[0028] Specifically, the preparation method of the attenuated Nocardia strain of yellowtail comprises the following steps:
[0029] S1. Construction of a homologous recombination vector: aseptically inoculate the preserved Nocardia spp. ZJ0503 strain into Brain Heart Infusion (BHI) solid medium (purchased from Guangdong Huanka...
Embodiment 2
[0047] This embodiment provides an attenuated strain of Nocardia japonica, which is a Nocardia japonica strain containing a deletion of the alanine dehydrogenase gene; the nucleotide of the alanine dehydrogenase gene The sequence is shown in the sequence listing SEQ ID NO: 1, wherein the nucleotide sequence of the upstream fragment of the alanine dehydrogenase gene is shown in the sequence listing SEQ ID NO: 2, and the downstream fragment of the alanine dehydrogenase gene The nucleotide sequence is shown in the sequence listing SEQ ID NO:3.
[0048] Specifically, the preparation method of the attenuated Nocardia strain of yellowtail comprises the following steps:
[0049] S1. Construction of a homologous recombination vector: aseptically inoculate the preserved Nocardia spp. ZJ0503 strain into Brain Heart Infusion (BHI) solid medium (purchased from Guangdong Huankai Microbial Technology Co., Ltd. ), cultured upside down at 26°C. After a single colony grows on the plate, pick...
Embodiment 3
[0055] This embodiment provides an attenuated strain of Nocardia japonica, which is a Nocardia japonica strain containing a deletion of the alanine dehydrogenase gene; the nucleotide of the alanine dehydrogenase gene The sequence is shown in the sequence listing SEQ ID NO: 1, wherein the nucleotide sequence of the upstream fragment of the alanine dehydrogenase gene is shown in the sequence listing SEQ ID NO: 2, and the downstream fragment of the alanine dehydrogenase gene The nucleotide sequence is shown in the sequence listing SEQ ID NO:3.
[0056] Specifically, the preparation method of the attenuated Nocardia strain of yellowtail comprises the following steps:
[0057] S1. Construction of a homologous recombination vector: aseptically inoculate the preserved Nocardia spp. ZJ0503 strain into Brain Heart Infusion (BHI) solid medium (purchased from Guangdong Huankai Microbial Technology Co., Ltd. ), cultured upside down at 30°C. When a single colony grows on the plate, pick ...
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