THE BACILLUS-SUBTILIS PHOAIV GENE - EFFECTS OF INVITRO INACTIVATION ON TOTAL ALKALINE-PHOSPHATASE PRODUCTION

被引:19
|
作者
KAPP, NV [1 ]
EDWARDS, CW [1 ]
CHESNUT, RS [1 ]
HULETT, FM [1 ]
机构
[1] UNIV ILLINOIS, DEPT BIOL SCI, MOLEC BIOL LAB, MC 067, POB 4348, CHICAGO, IL 60680 USA
关键词
Insertional mutagenesis; pho regulon; phosphate starvation; recombinant DNA;
D O I
10.1016/0378-1119(90)90346-S
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
A degenerative oligodeoxyribonucleotide probe deduced from the first 19 amino acids of the mature alkaline phosphatase IV (APase IV) protein was used to clone a DNA fragment internal to the coding region of the phoAIV gene of Bacillus subtilis. An insertional mutation was constructed in the phoAIV locus using the integrative plasmid, pJM103, containing the cloned DNA fragment. The strain with the interrupted phoAIV gene showed no detectable APase IV product on Western-blot analysis. The impact of the phoAIV interruption on total APase production in B. subtilis 168 was analyzed under both phosphate starvation and sporulation culturing conditions. The mutation in phoAIV reduced total APase-specific activity by 75% in phosphate-starved cells, and resulted in the elimination of a salt-extractable membrane APase, as well as the secreted APase IV. Analysis of this membrane APase indicated that it is a phoAIV gene product which is localized within the membrane fraction of the lysed cell and not secreted. There was no effect on the production of sporulation APase. The phoAIV::pJM103 insertion was mapped and determined to be located at approx. 73° on the B. subtilis 360° chromosome. © 1990.
引用
收藏
页码:95 / 100
页数:6
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