[Expression of phosphofructokinase gene from Escherichia coli K-12 in obligately autotrophic bacterium Acidithiobacillus thiooxidans].
A plasmid pSDK-1 containing the Escherichia coli phosphofructokinase-1 (EC 2.7.1. 11) gene (pfkA) was constructed and transferred into Acidithiobacillus thiooxidans Tt-Z2 by conjugation. The transfer frequency of plasmid from E. coli to Tt-Z2 was 2.6 x 10(-6). More than 68% of Tt-Z2 cells carried the recombinant plasmids after being cultured for 50 generations without selective pressure, which showed that pSDK-1 was maintained consistently in Tt-Z2. The pfkA gene from E. coli could be expressed in this obligately autotrophic bacterium but the enzyme activity (14 U/g was lower than that in E. coli (K-12: 86 U/g; DF1010 carrying plasmid pSDK-1: 97 U/g). In th presence of glucose, the Tt-Z2 transconjugant consumed glucose leading to a better growth yield.
Tian
Keli
K
Institute of Microbiology, Shandong University, Jinan 250100, China. tianzr@sdu.edu.cn
Lin
Jianqun
J
Liu
Xiangmei
X
Liu
Ying
Y
Zhang
Changkai
C
D013856:Acidithiobacillus thiooxidans / Q000235:genetics / Q000378:metabolism*; D052818:Autotrophic Processes; D003227:Conjugation, Genetic; D048168:Escherichia coli K12 / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D025221:Phosphofructokinases / Q000235:genetics* / Q000378:metabolism
D029968:Escherichia coli Proteins; D025221:Phosphofructokinases