[Cloning, characterization and expression in E. coli of the flagellin gene from Leptospira interrogans Serovar lai].
To obtain a large amount of recombinant endoflagellin protein of leptospsira, we have cloned the fla B gene from Leptospira interrogans serovar lai strain 017, which encodes the core flagellar protein. The entire open reading frame was amplified by polymerase chain reaction, which was digested with endonucleases Sal I and Cla I, then was orientationally cloned into the expression vector, pT7-7. The recombinant plasmid was transformed into Escherichia coli JM109 (DE3) for expression. After induction with IPTG, a 34 kd expression protein was detected by SDS-PAGE. The amount of recombinant protein was estimated as 11.8% of the whole bacterial lysate proteins. Probed with insert fragments, Southern blot analysis indicated that possibly two gene related to fla B were present in the genome of Leptospira interrogans serovar lai stain 017. Western blot analysis showed that the 34 kd expression protein reacting with rabbit polyclonal antiserum raised against leptospira endflagellin.
You
Z
Z
Research Unit of Leptospirosis, School of Basic Medical Sciences, Chengdu.
Dai
B
B
Fang
Z
Z
Li
S
S
Liu
J
J
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005408:Flagellin / Q000096:biosynthesis / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D007921:Leptospira interrogans / Q000235:genetics*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis
D011994:Recombinant Proteins; D005408:Flagellin