Expression vector of bacillus subtilis, recombinant strain and application thereof

Abstract

The present application relates to the field of microorganism and genetic engineering technology, and discloses an expression vector of bacillus subtilis, a recombinant strain and application.The expression vector contains the bglA gene of bacillus subtilis; the preservation number of the bacillus subtilis is GDMCC No:62938, and the preservation organization is Guangdong Microbial Culture Collection Center; the nucleotide sequence of the bglA gene of the bacillus subtilis is shown as SEQ ID NO:1.A new expression vector and recombinant strain are constructed, and recombinant expression and purification are realized in escherichia coli, and the recombinant protein can be used for subsequent enzymatic determination.The engineered escherichia coli expression system is highly mature in the aspects of host strain, promoter, induction mode and purification process, has the advantages of fast growth, convenient genetic operation and amplification culture, and can often obtain high expression, and is more suitable for rapid protein preparation and enzymatic verification of candidate genes.

Claims

1. An expression vector for Bacillus subtilis, characterized in that, The expression vector contains the bglA gene of Bacillus subtilis; The Bacillus subtilis has the following accession number: GDMCC No:62938, and the depository institution is Guangdong Provincial Center for Microbial Culture Collection. The nucleotide sequence of the bglA gene of Bacillus subtilis is shown in SEQ ID NO:

1.

2. The expression vector according to claim 1, characterized in that, The expression vector was constructed by inserting the bglA gene of Bacillus subtilis into plasmid pET-32a(+).

3. The expression vector according to claim 2, characterized in that, The bglA gene of Bacillus subtilis was inserted into the Sac I and Xho I sites of plasmid pET-32a(+) to obtain the bglA-pET32α(+) expression vector.

4. A recombinant strain comprising the expression vector according to any one of claims 1-3.

5. The recombinant strain according to claim 4, characterized in that, The recombinant strain has the following accession number: GDMCCNo:67748, and the depository is Guangdong Provincial Center for Microbial Culture Collection.

6. The recombinant strain according to claim 4, characterized in that, The bglA gene of Bacillus subtilis was inserted into the Sac I and Xho I sites of plasmid pET-32a(+) to form the bglA-pET32α(+) expression vector carrying the bglA gene of Bacillus subtilis; the bglA-pET32α(+) expression vector was transformed into E. coli BL21(DE3) and cultured to form a recombinant strain.

7. The use of the recombinant strain or its agent according to any one of claims 4-6 in the degradation of glucosinolates.

8. The use of the recombinant strain or its inoculum according to any one of claims 4-6 in feed fermentation.

9. The application according to claim 8, characterized in that, The feed includes rapeseed meal.

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