Cloning and sequence analysis of the gene encoding LipL32 of Leptospira interrogans serovar Sejroe.
Leptospira, a member of the order Spirochaetales, is the causative agent of leptospirosis, an important zoonosis encountered worldwide. The Leptospira interrogans serovar Sejroe was grown in EMJH medium and its DNA was isolated using standard techniques. The LipL32 gene was amplified using the reported primer of Kirschneri of LipL32. The amplified product was found to comprise 756 base pairs. This amplified gene fragment of LipL32 lipoprotein was cloned in E. coli (DH5 alpha) cells using pDrive plasmid as a vector. The recombinant cells were selected on LB agar medium containing ampicillin, X-gal and isopropyl-beta-D-thiogalactopyranoside. Plasmid was extracted from the recombinant white colonies, and restriction endonuclease (RE) analysis was carried out using PstI and SalI. On partial sequence analysis, the product exhibited 756 base pairs, corresponding to 251 amino acids. The cloned gene could be further used for expression of recombinant protein for serodiagnosis of leptospirosis.
Amutha
R
R
Division of Bacteriology and Mycology, Indian Veterinary Research Institute, Izatnagar, India.
Chaudhury
P
P
Garg
Amar P
AP
Vasan
P
P
Cheema
P S
PS
Srivastava
S K
SK
D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D014644:Genetic Variation; D007921:Leptospira interrogans / Q000145:classification* / Q000235:genetics*; D008074:Lipoproteins / Q000235:genetics*; D010802:Phylogeny
D001425:Bacterial Outer Membrane Proteins; C406589:LipL32 protein, Leptospira; D008074:Lipoproteins