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US 2004/0009556 A1Published application
Natural promoters for gene expression and metabolic monitoring in bacillus species
OwnerLaura A. Bedzyk, Tao Wang +1inventors
InventorsLaura A. Bedzyk, Tao Wang, Rick W. Ye
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Abstract
Genes have been identified in the Bacillus genome that are responsive to various metabolic conditions and growth cycle changes. The new responsiveness of these genes allows for their use in regulated gene expression in Bacillus sp. and for the monitoring of bioreactor health.
Claims28
A method for the expression of a coding region of interest in a Bacillus sp comprising:
- a) providing a transformed Bacillus sp cell having a chimeric gene comprising a nucleic acid fragment comprising the promoter region of a Bacillus gene operably linked to a coding region of interest expressible in a Bacillus sp, wherein the nucleic acid fragment comprising the promoter region of a Bacillus gene is selected from the group consisting of narGHJI, csn, yncM, yvyD, yvaWXY, ydjL, sunA, and yolIJK and homologues thereof;
- and
- b) growing the transformed Bacillus sp cell of step (a) in the absence of oxygen wherein the chimeric gene of step (a) is expressed.
A method for the expression of a coding region of interest in a Bacillus sp comprising:
- a) providing a transformed Bacillus sp cell having a chimeric gene comprising a nucleic acid fragmnent comprising the promoter region of a Bacillus gene operably linked to a coding region of interest expressible in a Bacillus sp, wherein the nucleic acid fragment comprising the promoter region of a Bacillus gene is selected from the group consisting of narGHJI, csn, yncM, yvyD, yvaWXY, ydjL, sunA, and yolIJK and homologues thereof;
- b) growing the transformed Bacillus sp. cell of step (a) in the presence of oxygen whereby the cell density is increased;
- and
- c) removing oxygen form the transformed Bacillus sp. cell or step (b) whereby the chimeric gene is expressed.
1 more element
A method for the expression of a coding region of interest in a Bacillus sp comprising:
- a) providing a transformed Bacillus sp cell having a chimeric gene comprising a nucleic acid fragment comprising the promoter region of a Bacillus gene operably linked to a coding region of interest expressible in a Bacillus sp, wherein the nucleic acid fragment comprising the promoter region of a Bacillus gene is selected from the group consisting of feuABC, ykuNOP, and dhbABC, and homologues thereof;
- and
- b) growing the transformed Bacillus sp cell of step (a) in the absence of oxygen and in the presence of nitrite wherein the chimeric gene of step (a) is expressed.
A method for the expression of a coding region of interest in a Bacillus sp comprising:
- a) providing a transformed Bacillus sp cell having a chimeric gene comprising a nucleic acid fragment comprising the promoter region of a Bacillus gene operably linked to a coding region of interest expressible in a Bacillus sp, wherein the nucleic acid fragment comprising the promoter region of a Bacillus gene is selected from the group consisting of ycgMN, dhaS rapF, rapG, rapH, rapK, yqhIJ, yveKLMNOPQST, yhfSTUV, csn, yncM, yvyD, yvaWXY, ydjL, sunA, and yolIJK, and homologues thereof;
- and
- b) growing the transformed Bacillus sp cell of step (a) in the presence of oxygen until the cell reaches about T0 of the stationary phase wherein the chimeric gene of step (a) is expressed.
A method for the expression of a coding region of interest in a Bacillus sp comprising:
- a) providing a transformed Bacillus sp cell having a chimeric gene comprising a nucleic acid fragment comprising the promoter region of a Bacillus gene operably linked to a coding region of interest expressible in a Bacillus sp, wherein the nucleic acid fragment comprising the promoter region of a Bacillus gene is selected from the group consisting of acoABCL, and glvAC, and homologues thereof;
- and
- b) growing the transformed Bacillus sp cell of step (a) in the presence of oxygen until the cell reaches about T1 of the stationary phase wherein the chimeric gene of step (a) is expressed.
A method for the expression of a coding region of interest in a Bacillus sp comprising:
- a) providing a transformed Bacillus sp cell having a chimeric gene comprising a nucleic acid fragment comprising the promoter region of a Bacillus gene operably linked to a coding region of interest expressible in a Bacillus sp, wherein the nucleic acid fragment comprising the promoter region of a Bacillus gene is selected from the group consisting of yxjCDEF, yngEFGHI, yjmCDEFG, ykfABCD, and yodOPRST;
- and homologues thereof;
- and
- b) growing the transformed Bacillus sp cell of step (a) in the presence of oxygen until the cell reaches about T3 of the stationary phase wherein the chimeric gene of step (a) is expressed.
1 more element
A method for monitoring the state of the cell metabolism of a Bacillus sp. culture comprising:
- a) providing a culture of actively growing Bacillus sp. cells;
- and
- b) measuring the expression levels of a pool of genes isolated from the Bacillus cells of step (a), the pool of genes comprising narGHJI, feuABC, ykuNOP, dhbABC, ydjL, sunA, yolIJK, csn, yncM, yvyD, yvaWXY, yhfRSTUV, yveKLMNOPQST, dhaS, rapF, rapG, rapH, rapK, ycgMN, yqhIJ, glvAC, acoABCL, yxjCDEF, yngEFGHI yjmCDEFG, ykfABCD, yodOPRST, alsT, and yxeKLMN, and homologues thereof.
Dependencies are inferred from the claim text.