1.Construction of a temperature inducible shuttle expression vector and its application in Streptomyces.
Mei-Feng TAO ; Xiu-Fen ZHOU ; Tobias KIESER ; Zi-Xin DENG
Chinese Journal of Biotechnology 2002;18(4):420-423
pHZ1080, an E. coli-Streptomyces shuttle expression vector was constructed in order to explore the utilization of lambda phage regulated expression elements in Streptomyces. A 2.7 kb polyketide synthase (PKS) gene from Streptomyces sp. FR-008 was inserted into downstream of lambda phage promoter (PR) to give the shuttle plasmid, pHZ1067. The PKS protein was expressed in Streptomyces lividans carrying pHZ1067 in a heat-dependent manner, as it did in E. coli. The PKS protein expressed in both hosts with same molecular weight was detected by SDS-PAGE and Western-blot. The successful heat-induced expression of PKS suggested that pHZ1080 was useful and convenient for heat-induced expression of heterologous genes in both E. coli and Streptomyces.
Amino Acid Sequence
;
Base Sequence
;
Blotting, Western
;
Cloning, Molecular
;
Genetic Vectors
;
genetics
;
Molecular Sequence Data
;
Multienzyme Complexes
;
genetics
;
metabolism
;
Recombinant Proteins
;
genetics
;
metabolism
;
Streptomyces
;
enzymology
;
genetics
;
Temperature