Expression vector for pseudonocardia autotrophica

1. An expression vector, including a replication initiation region derived from Pseudonocardia autotrophica , a multicloning site for introducing an exogenous gene, an exogenous gene introduced into the multicloning site, a promoter, a terminator, and a defective selection marker, which autonomously replicates in cells of Pseudonocardia autotrophica to enable expression of the exogenous gene introduced, wherein the replication initiation region comprises a base sequence represented by SEQ ID NO: 49, a base sequence complementary to the entire base sequence represented by SEQ ID NO: 49, a base sequence having 90% or more homology to SEQ ID NO: 49, or a base sequence complementary to the entire base sequence having 90% or more homology to SEQ ID NO: 49.

2. The expression vector according to claim 1 , in which the promoter is induced by acetone to express the exogenous gene.

3. The expression vector according to claim 2 , in which the promoter region comprises a base sequence represented by SEQ ID NO: 26 or a base sequence having 90% or more homology to the above base sequence or a complementary sequence thereof.

4. The expression vector according to claim 1 , further including a replication initiation region derived from Escherichia coli , and being autonomously replicable in both Pseudonocardia autotrophica and Escherichia coli , and can be used as a shuttle vector.

5. The expression vector according to claim 4 , further comprising an oriT region and which can perform transformation by conjugation of Escherichia coli S17-1 and Pseudonocardia autotrophica.

6. The expression vector according to claim 1 , in which the exogenous gene is a gene encoding vitamin D hydroxylase or a gene encoding compactin hydroxylase.

7. A transformant of Pseudonocardia autotrophica having introduced thereinto the expression vector according to claim 1 .

8. A method of producing a protein comprising: introducing the expression vector according to claim 1 into Pseudonocardia autotrophica to be transformed; and expressing the exogenous gene in the resultant transformant to produce a protein.

9. A method of producing 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, or an active form of vitamin D including: transforming Pseudonocardia autotrophica with the expression vector according to claim 1 , having introduced thereinto a vitamin D hydroxylase gene as an exogenous gene; inducing expression of vitamin D hydroxylase from the resultant transformant; and culturing the transformant with vitamin D group, thereby producing 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, or an active form of the vitamin D group.

10. The method of producing an active form of vitamin D according to claim 9 , in which the active form of vitamin D is 1α,25-dihydroxyvitamin D3.

11. A method of producing pravastatin including: transforming Pseudonocardia autotrophica with the expression vector according to claim 1 , having introduced thereinto a compactin hydroxylase gene as an exogenous gene; inducing expression of compactin hydroxylase from the resultant transformant; and culturing the transformant with compactin, thereby producing pravastatin from compactin.

12. The expression vector according to claim 1 , wherein the replication initiation region is selected from the group consisting of rep1 and rep5.

13. The expression vector according to claim 1 , wherein the promoter is selected from the group consisting of an acetone-inducible promoter, a thiostrepton-inducible promoter, and an ermE promoter.

14. The expression vector according to claim 1 , wherein the vector is selected from the group consisting of pTAOR3-vdh, pTAOR4-For-boxAB and pTAOR4-Rev-boxAB.