CLONING AND EXPRESSION OF RECOMBINANT LIPL32 ANTIGEN FROM LEPTOSPIRA INTERROGANS IN PROKARYOTIC SYSTEM

سال انتشار: 1397
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 388

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شناسه ملی سند علمی:

MEDISM19_655

تاریخ نمایه سازی: 13 مهر 1397

چکیده مقاله:

Background and Aim:Leptospirosis,caused by infection with pathogenic leptospira species is recognized as the most widespread zoonosis with a global distribution. Lipl32 is an immunogenic outer membrance protein found in pathogenic Leptospira species. The protein expressed by lipl32 gene may be used in diagnostic method and also can be a good candidate for recombinant vaccine against leptospirosis. The aim of this study was Cloning and expression of recombinant LipL32 antigen from Leptospira interrogans in prokaryotic systemMethods:LipL32 was clonned in E. coli strain BL21 using pET32 + plasmid as a vector and induced with 25 µM IPTG at 22 °C for 16 hours. The bacterial pellet was obtained by centrifugation (5,000 rpm). The resuspended cells were disrupted by sonication. The cell lysate was centrifuged at 15,000 rpm at 4 °C for 20 min. Samples were solubilized in sample buffer plus mercaptoenthaol , Proteins were separated on 10% sodium dodecyl sulfate (SDS) and followed by Coomassie Brilliant Blue stainingResults:The protein was successfully expressed in E.coli BL21 and purified. SDS-PAGE results showed that the full-length 38kD protein was induced by 25 µM IPTGConclusion:Leptospirosis is considered as a reemerging infectious disease, not only for the increase in its incidence during the past recent years but also for the increased severity of the illness. The cloned gene could be further used for expression of recombinant protein for serodiagnosis and leading candidate vaccine antigens of leptospirosis.

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نویسندگان

Nina Bakhshandeh

Department of Biology,Science and Research Branch,Islamic Azad university,Tehran,Iran

Majid Tebianian

Razi Vaccine & Serum Research Institute,karaj,iran

Pejvak Khaki

Razi Vaccine & Serum Research Institute,karaj,iran

Majid Esmaeilzad

Razi Vaccine & Serum Research Institute,karaj,iran