Expression and characterization of recombinant leptospiral outer membrane protein LipL32 from Leptospira interrogans serovar autumnalis.
Leptospira interrogans serovar autumnalis, a causative agent of leptospirosis in Thailand, was isolated from a patient for DNA extraction and amplification of LipL32 gene by polymerase chain reaction (PCR). The 782 bp PCR product was obtained, which was inserted into pAE plasmid with polyhistidine (His6 tag) to construct pAE-LipL32. This recombinant plasmid was transfected into E. coli BL21 (DE3). His6-LipL32 was purified by Ni-NTA affinity chromatography. The recombinant protein was used as antigen for testing with sera from leptospirosis and syphilis patients by dot-ELISA technique. It reacted positively with leptospirosis patient sera and negatively with syphilis and healthy sera.
Boonsathorn
Naphatsawan
N
Medical Biotechnology Center, Department of Medical Sciences, Ministry of Public Health, Nonthaburi, Thailand. naphatsawan@dmsc.moph.go.th
Konghom
Ganokrot
G
Mongkolsiri
Kaveewan
K
Jirapongwattana
Chanin
C
Balachandra
Kruavon
K
Naigowit
Pimjai
P
Sawanpanyalert
Pathom
P