expression and purification of neurotoxin-associated protein ha-33/a from clostridium botulinum and evaluation of its antigenicity
نویسندگان
چکیده
background: botulinum neurotoxin (bont) complexes consist of neurotoxin and neurotoxin-associated proteins. hemagglutinin-33 (ha-33) is a member of bont type a (bont/a) complex. considering the protective role of ha-33 in preservation of bont/a in gastrointestinal harsh conditions and also its adjuvant role, recombinant production of this protein is favorable. thus in this study, ha-33 was expressed and purified, and subsequently its antigenicity in mice was studied. methods: initially, ha-33 gene sequence of clostridium botulinum serotype a was adopted from genbank. the gene sequence was optimized and synthesized in pet28a (+) vector. e. coli bl21 (de3) strain was transformed by the recombinant vector and the expression of ha-33 was optimized at 37°c and 5 h induction time. results: the recombinant protein was purified by nickel nitrilotriacetic acid agarose affinity chromatography and confirmed by immunoblotting. enzyme linked immunoassay showed a high titer antibody production in mice. conclusion: the results indicated a highly expressed and purified recombinant protein, which is able to evoke high antibody titers in mice.
منابع مشابه
Expression and Purification of Neurotoxin-Associated Protein HA-33/A from Clostridium botulinum and Evaluation of Its Antigenicity
Background: Botulinum neurotoxin (BoNT) complexes consist of neurotoxin and neurotoxin-associated proteins. Hemagglutinin-33 (HA-33) is a member of BoNT type A (BoNT/A) complex. Considering the protective role of HA-33 in preservation of BoNT/A in gastrointestinal harsh conditions and also its adjuvant role, recombinant production of this protein is favorable. Thus in this study, HA-33 was expr...
متن کاملExpression and purification of neurotoxin-associated protein HA-33/A from Clostridium botulinum and evaluation of its antigenicity.
BACKGROUND Botulinum neurotoxin (BoNT) complexes consist of neurotoxin and neurotoxin-associated proteins. Hemagglutinin-33 (HA-33) is a member of BoNT type A (BoNT/A) complex. Considering the protective role of HA-33 in preservation of BoNT/A in gastrointestinal harsh conditions and also its adjuvant role, recombinant production of this protein is favorable. Thus in this study, HA-33 was expre...
متن کاملCloning, Expression and Purification of Clostridium botulinum Neurotoxin Type E Binding Domain
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extraction and acetylation of purified trypsin from bovin pancreas and study of some its physico-chemical properties
آنزیم تریپسین در شرایط قلیایی ناپایدار می باشد .و فعالیت پروتئولیتیکی تریپسین منجربه خود هضمی آن در جایگاههای خاصی می گردد. بنابر این آنزیمی با ناپایداری بالا محسوب میگردد. در سالهای اخیر موفق شدند که با ایجاد تغیرات شیمیایی با اضافه کردن فلزات خاص ، کلسیم و یا عمل استیلاسیون منجر به افزایش پایداری آنزیم تریپسین گردند. مطالعات در حال حاضر نشان می دهد که تریپسین استیله شده فعالیت آنزیمی خود را ...
15 صفحه اولPurification and Characterization of Botulinum Neurotoxin FA from a Genetically Modified Clostridium botulinum Strain
Botulinum neurotoxins (BoNTs), produced by neurotoxigenic clostridial species, are the cause of the severe disease botulism in humans and animals. Early research on BoNTs has led to their classification into seven serotypes (serotypes A to G) based upon the selective neutralization of their toxicity in mice by homologous antibodies. Recently, a report of a potential eighth serotype of BoNT, des...
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عنوان ژورنال:
iranian biomedical journalجلد ۱۷، شماره ۴، صفحات ۱۶۵-۱۷۰
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