amplification and cloning of a gene encoding a 41 kda outer membrane protein (lipl41) of leptospira interrogans serovar canicola

نویسندگان

pejvak khaki

national reference laboratory for leptospira, department of microbiology, razi vaccine & serum research institute, karaj, iran soheila moradi bidhendi

national reference laboratory for leptospira, department of microbiology, razi vaccine & serum research institute, karaj, iran yung-fu chang

department of population, medicine and diagnostic sciences, college of veterinary medicine, cornell university, ithaca, ny, usa maryam sadat soltani

national reference laboratory for leptospira, department of microbiology, razi vaccine & serum research institute, karaj, iran kayvan tadaion

چکیده

background: leptospirosis has been recognized as an important reemerging infectious disease caused by pathogenic leptospira spp. a major challenge of this disease is the application of a basic research to improve diagnostic method. outer membrane proteins of leptospira are potential candidates that could be useful in diagnosis. among them the lipl41 is an immunogenic protein which is present only in pathogenic serovars. in order to evaluate genetic conservation of the lipl41 gene, we cloned and sequenced this gene from leptospira interrogans serovar canicola. materials and methods: following the dna extraction from the serovar, the lipl41 gene was amplified and cloned into ptz57r/t vector and transformed into the competent e. coli (top10). recombinant clones were confirmed by colony pcr and dna sequencing. the related sequences were then analyzed and compared with the sequences in the genbank database. results: pcr amplification of the lipl41 gene resulted in a 1065 bp pcr product. the pcr based on the lipl41 gene detected all the pathogenic reference serovars of the tested leptospira spp. it was revealed that in iran the homology of the lipl41 gene between vaccinal and clinical serovars of canicola was 100%. it also showed >95.9% homology with other pathogenic serovars in genbank database, which indicates genetic conservation of this gene. conclusion: because of the conservation of lipl41 gene among different strains of leptospira and its exclusive presence in leptospira, it was revealed that the cloned gene could be further used as a good candidate for developing diagnostic methods such as elisa and as positive control in diagnostic pcr.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Immunoreactive outer membrane proteins of Leptospira interrogans serovar Canicola strain Hond Utrecht IV.

BACKGROUND & OBJECTIVES Leptospirosis is a severe and complex zoonotic disease prevalent in many countries including India. Current leptospiral research is focussed on the identification of the outer membrane proteins (OMPs) of the organism that could be used in developing diagnostic assays for leptospirosis. METHODS The Leptospira interrogans serovar Canicola was grown in EMJH medium and the...

متن کامل

Evaluation of recombinant Leptospira interrogans serovar canicola outer membrane proteins as diagnostic antigen.

Leptospirosis of animals and man is an important rereported in recent years are LipL32, LipL41 and OmpL1; they emerging infectious disease worldwide. In domestic animals are expressed in-vivo during infection. LipL32 and LipL41 it is an important cause of abortion, stillbirth, infertility, are important and surface-exposed leptospiral outer membrane decreased milk production and death. In dairy...

متن کامل

Vaccination with leptospiral outer membrane lipoprotein LipL32 reduces kidney invasion of Leptospira interrogans serovar canicola in hamsters.

The Leptospira interrogans vaccines currently available are serovar specific and require regular booster immunizations to maintain protection of the host. In addition, a hamster challenge batch potency test is necessary to evaluate these vaccines prior to market release, requiring the use of a large number of animals, which is ethically and financially undesirable. Our previous work showed that...

متن کامل

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 (DE...

متن کامل

LipL41, a Hemin Binding Protein from Leptospira santarosai serovar Shermani

Leptospirosis is one of the most widespread zoonotic diseases in the world. It is caused by the pathogen Leptospira that results in multiple-organ failure, in particular of the kidney. Outer membrane lipoprotein is the suspected virulence factor of Leptospira. In Leptospira spp LipL41 is one major lipoprotein and is highly conserved. Previous study suggests that LipL41 bears hemin-binding abili...

متن کامل

Evolutionary Implication of Outer Membrane Lipoprotein-Encoding Genes ompL1, lipL32 and lipL41 of Pathogenic Leptospira Species

Leptospirosis is recognized as the most widespread zoonosis with a global distribution. In this study, the antigenic variation in Leptospira interrogans and Leptospira borgpetersenii isolated from human urine and field rat kidney was preliminarily confirmed by microscopic agglutination test using monoclonal antibodies, and was further subjected to amplification and identification of outer membr...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
infection, epidemiology and medicine

جلد ۲، شماره ۳، صفحات ۵-۷

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023