[Heterologous genes expression on Escherichia coli chromosome lac operon using Red recombination].
To achieve efficient and stable expression of heterologous exogenetic protein or antigen in E. coli chromosome, the luciferase report gene was knocked in lacZ site of chromosome lac operon by using Red recombination system and selection-counterselection kan/sacB technology. The quantitative analysis of exogenous gene expression indicated that the target gene could be efficiently expressed at lacZ site of lac operon. The results confirmed the efficient screening and stable expression of heterologous protein or antigen on chromosome by using the recombinant engineering technique. This study demonstrated that the chromosome could be used as a vector for heterologous protein or antigen and the stable expression of exogenous gene on E. coli chromosome had no side effect on the bacterial growth and propagation.
Li
Shanhu
S
Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100850, China.
Shi
Qingguo
Q
Huang
Cuifen
C
Zhou
Jianguang
J
D010582:Bacteriophage lambda / Q000235:genetics; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D055879:Gene Knock-In Techniques / Q000379:methods; D017930:Genes, Reporter / Q000235:genetics; D007763:Lac Operon / Q000235:genetics*; D008156:Luciferases / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics*
D008156:Luciferases